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1, 6-glucans; and mannoproteins in broiler chicken diets
Use of yeast cell walls; β-1, 3/1, 6-glucans; and mannoproteins in broiler chicken diets R. Morales-López,*1 E. Auclair,† F. García,‡ E. Esteve-Garcia,* and J. Brufau* *Institute of Research, Food Technology and Agriculture, Department of Animal Nutrition, Centre Mas de Bover, Carr. Reus-Morell, km 3.8, 43120 Constantí (Tarragona), Spain; †Lesaffre Feed Additives, Rue du Haut Touquet 1, 59520 Marquette-Lez-Lille, France; and ‡Saf-Agri, México, km. 57.5, Carr. México-Toluca, C.P. 50200 Toluca, Estate of Mexico, Mexico
Key words: mannoprotein complex, β-1, 3/1, 6-glucan, yeast cell wall, broiler chicken, villi height 2009 Poultry Science 88:601–607 doi:10.3382/ps.2008-00298 of mannan sugar polymers), 15 to 30% proteins, 5 to 20% lipids, and no more than 5% of chitin (AguilarUscanga and François, 2003; EURASYP, 2007). Most of the protein is linked to the mannanoligosaccharides (MOS) and is referred to as the mannoprotein complex (MP). In the digestive tract of animals, MOS present in YCW could act as high-affinity ligands, with the potential benefit of offering a competitive binding site for pathogenic bacteria mannose-specific type-1 fimbriae (Spring et al., 2000). The β-1, 3/1, 6-glucans also present in YCW should be recognized as an immune modulator substance in animals and humans (Abel and Czop, 1992; Raa, 2003); thus, dietary YCW might exert some benefits on the immune system of intestinal mucosa (Ferket et al., 2002). The importance of β-1, 3/1, 6-glucans and MP-purified fractions on the mode of action for YCW additives has not been studied in poultry. Thus, the aim of these studies was to evaluate the effect of YCW obtained from S. cerevisiae of
INTRODUCTION Saccharomyces cerevisiae yeast cell wall (YCW) components have been used in animal feeding since the last decades (Hooge, 2004; Rosen, 2007). Their inclusion in broiler diets has resulted in improvements of animal productivity, which was attributed to physiological effects on intestinal digestive mucosa (Santin et al., 2001; Zhang et al., 2005; Baurhoo et al., 2007). However, the mode of action of YCW products in broiler chicken diets is not well understood and the characteristics of YCW products have been poorly defined. Typically, commercial YCW are composed of 30 to 60% polysaccharides (15 to 30% of β-1, 3/1, 6-glucan and 15 to 30% ©2009 Poultry Science Association Inc. Received July 22, 2008. Accepted October 27, 2008. 1 Corresponding author: [email protected]
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BG treatments. In experiment 2, there was a group-fed basal diet and 3 additional experimental groups receiving the basal diet supplemented with, respectively: 500 mg of YCW/kg of feed, 190 mg of MP/kg of feed, and 227 mg of BG/kg of feed. At 42 d of age, no difference in broiler growth rates was observed. Samples of the jejunum were collected at 21 d of age to determine villus height. Significantly higher villus height was observed in YCW, MP, and BG groups compared with that of the control group. The relative percentage of liver weight (P ≤ 0.01) was lower in broilers fed YCW than in broilers fed the control diet, but no differences were observed in respect to chickens fed BG. Data of these studies suggested that the changes in thymus and liver relative weights and villus morphology of broilers were induced with the same intensity by the use of complete YCW, MP + BG, and BG supplements.
ABSTRACT Two experiments were carried out to evaluate the effect of dietary addition of yeast cell wall (YCW); β-1, 3/1, 6-glucan (BG); and mannoprotein complex (MP) purified fractions in broilers. In experiment 1, there was a control diet and 5 experimental diets containing, respectively: 10 mg of avilamycin (AVI)/ kg of feed, 500 mg of YCW/kg of feed, 95 mg of MP/ kg of feed, 145 mg of BG/kg of feed, and 95 mg of MP plus 145 mg of BG/kg of feed. All birds were vaccinated via drinking water against Newcastle disease virus at 9 d of age. At 42 d, chickens fed AVI, YCW, MP + BG, and BG diets had similar BW not significantly different from chickens fed the control diet. The antibody response of Newcastle disease virus vaccine was not affected by any experimental treatment. Broilers fed MP + BG diet had greater thymus weights (P ≤ 0.05), as a percentage of BW than those from the control and AVI treatments, but similar with respect to YCW and
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the yeast extract industry and their MP and β-1, 3/1, 6-glucans-purified fractions supplemented in the diets of broiler chickens.
MATERIALS AND METHODS Animal Housing
Diets The YCW efficacy was tested on wheat-based diets in mash form without any antibiotic growth promoters (AGP) and coccidiostats (Table 1). There were no exogenous enzymes used for wheat, barley, and rye cereals because they might mask the effects of yeast additives. Diets were formulated to meet or exceed NRC (1994) requirements. In experiment 1, a single diet was used throughout the 42 d of trial to avoid adaptation to the change of diet. In experiment 2, starter and grower diets were used from 0 to 21 and from 21 to 42 d, respectively (Table 1). The analytical composition of the main ingredients and feeds was determined using standard methods (AOAC, 1990) including DM (method 934.01), CP (method 968.06; protein-nitrogen determination, LECO FP-528, St. Joseph, MI), crude fat (method 920.39), and gross energy (DIN, 1977) using an adiabatic calorimeter bomb (Calorimeter C-4000 A, IKA Analysentrchnik GmbH, Heitersheim, Germany). In experiment 1, experimental YCW contained 19% mannose and 29% of β-1, 3/1, 6-glucans. In experiment 2, experimental YCW contained 21% of mannose and 25% of β-1, 3/1, 6-glucans; purified MP contained 55% mannose; and purified β-glucans (BG) contained 55% of β-1, 3/1, 6-glucans in β-1, 3 main forms. The determination of glucans and mannan polysaccharides of YCW, MP, and BG was conducted at Bio-Springer (Maisons-Alfort Cedex, France) by using the EURASYP (2007) methodology. In both experiments, chickens were weighed at 1, 21, and 42 d of age. The BW, daily weight gain, daily feed intake, and feed conversion ratio (FCR) were calculated for each period and for the overall experiment.
Experiment 1 evaluated the effect of YCW from the yeast extract industry (Lesaffre Feed Additives, Marquette-Lez-Lille, France) and their main polysaccharide components such as MP- and BG-purified fractions on growth performance parameters, antibody response against Newcastle disease virus (NDV) vaccine, and lymphoid organs (spleen, thymus, and bursa of Fabricius) relative weights (RW). A total of 828 broilers chickens were used. There was a control diet and 5 experimental diets containing, respectively: 10 mg of avilamycin (AVI)/kg of feed, 500 mg of YCW/kg of feed, 95 mg of MP/kg of feed, 145 mg of BG/kg of feed, and 95 mg of MP plus 145 mg of BG/kg of feed. Each treatment was replicated 6 times of 23 birds. At 9 d of age, all birds were vaccinated through drinking water with an attenuated NDV vaccine (“La Sota” strain). At 23 and 36 d, 3 chickens per pen (or 18 per treatment) were bled through the brachial vein to measure antibody titters against NDV by the hemagglutination inhibition test (Takahashi et al., 1994). At 36 d of age, 8 chickens from each treatment were selected at random and were killed according to protocols accepted by the Ethical Commission of Institute of Research, Food Technology and Agriculture (Catalonia, Spain). The spleen, bursa of Fabricius, and thymus (all right and left lobules) were excised and weighed. Afterwards, adherent fat from these organs was removed and their RW expressed as a percentage of live BW were calculated: RW = organ weight × 100/chicken BW.
Experiment 2 Experiment 2 evaluated the effect of YCW from the yeast extract industry, MP, and BG on growth performance, villus height of jejunal mucosa, and RW of digestive organs (intestine, pancreas, and liver). A total of 800 broiler chicks were used, and there was a control diet and 3 experimental diets containing, respectively: 500 mg of YCW/kg of feed, 190 mg of MP/kg of feed, and 227 mg of BG/kg of feed. Each treatment included 5 replicates of 40 chickens. At 21 d of age, 10 chickens from each treatment were killed. The complete intestine (from the gizzard-duodenal junction to rectum-cloacal junction), pancreas, and liver were excised and weighed. Afterward, adherent fat from these organs was removed and their RW were calculated. From the same killed chicks, 10 samples from each experimental treatment were collected from the proximal jejunum (5 cm) 2 cm following the end of the duodenal loop. Samples from jejunum were fixed in Carnoy’s solution (60% absolute ethanol + 30% chlorophorm + 10% glacial acetic acid-based solution), dehydrated (70% alcohol, 60 min; 96% alcohol, 45 min and then 60 min; isopropylic alcohol, 60 min), embedded in paraffin, and 4-μm sections were cut. Then, sections
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Two experiments were conducted using commercial 1-d-old Ross 308 male broiler chicks. Chickens were placed in 36 one-square meter cages in the first experiment (experiment 1) and in 20 floor pens of 4 square meters with wood shavings as litter in the second experiment (experiment 2). There were 6 and 5 blocks in experiment 1 and 2, respectively, corresponding to location in the experimental house, and treatments were randomly distributed within each block. The experimental house was provided with forced ventilation, artificial light, and gas heating. The temperature inside the house on arrival was 33 to 35°C and was decreased by 3°C each week until 22°C. The lighting program was 23 h of light the first 4 d, 20 h until 10 d, and 18 afterwards. Feed and water were provided for ad libitum consumption throughout the 42 d of experimentation.
Experiment 1
YEAST CELL WALLS; β-1, 3/1, 6-GLUCANS; AND MANNOPROTEINS
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Table 1. Ingredient composition (g/kg) and calculated nutrient analysis of experimental diets Experiment 1 Item
Single diet
Starter diet
Grower diet
437.2 150.0 50.0 185.5 109.7 30.0 11.81 14.51 2.61 1.15 — 3.00 0.05 4.00 0.50 1,000.0
379.8 124.9 50.0 232.3 120.0 48.0 11.90 17.10 2.70 2.20 0.80 3.20 1.10 4.00 2.00 1,000.0
519.6 63.2 25.0 152.6 140.0 58.0 10.30 16.00 2.10 2.70 0.70 3.20 0.60 4.00 2.00 1,000.0
3,000 20.0 1.10 0.55 0.90 0.72 0.90 0.60 0.40
3,000 22.0 1.27 0.58 0.94 0.85 1.00 0.68 0.45
3,150 20.0 1.15 0.50 0.83 0.76 0.90 0.64 0.42
20.3 6.2 89.7
22.6 8.3 89.5
19.3 9.2 89.7
1 Provided per kilogram of feed: vitamin A, 12,000 IU; vitamin D3, 2,400 IU; vitamin E, 30 mg; vitamin K3, 3 mg; thiamine, 2.2 mg; riboflavin, 8 mg; pyridoxine, 5 mg; cyanocobalamin, 11 μg; folic acid, 1.5 mg; biotin, 150 μg; calcium pantothenate, 25 mg; nicotinic acid, 65 mg; Mn (manganese sulfate), 60 mg; Zn (zinc oxide), 40 mg; I (potassium iodate), 0.33 mg; Fe (ferrous sulfate), 80 mg; Cu (copper sulfate), 8 mg; Se (sodium selenite), 0.15 mg; ethoxyquin, 150 mg. 2 The experimental yeast fractions: 500 g of yeast cell walls (YCW), 95 g of mannoprotein (MP), and 145 g of β-glucans (BG) in experiment 1 and 500 g of YCW, 190 g of MP, and 227 g of BG in experiment 2 were included per ton of experimental feed to expenses of corn starch.
were stained with Luxol Fast Blue/PAS stain (Polysciences Inc., Ajalvir-Madrid, Spain) to determine the villus height in at least 3 villi per jejunal sample.
Statistical Analysis Experiment 1 was analyzed by 2-way ANOVA as a randomized complete block design with 6 blocks corresponding to location within the house and 6 dietary treatments; experiment 2 was analyzed by the same statistical design as experiment 1 but with 5 blocks and 4 dietary treatments (SAS Institute, 1996). Differences between treatments were established by Tukey’s test at P ≤ 0.05 probability. The data expressed as percentages such as mortality were transformed to arcsine values.
RESULTS Experiment 1 Experimental diets did not affect growth performance parameters at different periods and cumulatively ex-
cept for FCR from 0 to 21 d (Table 2). The FCR was impaired (P ≤ 0.01) in chickens fed YCW and MP + BG compared with those fed the control diet, AVI, and BG. At 38 d, chickens fed MP + BG fractions had significantly heavier thymus RW (P ≤ 0.05) with respect to those fed the control and the AVI diets, and similar than those chicks fed YCW and BG (Table 3), and the bursa of Fabricius and spleen RW were not affected by treatment (Table 3). At 23 and 36 d, the antibody response of NDV vaccine did not show changes due to the use of experimental diet.
Experiment 2 Experimental diets did not affect growth performance parameters at different periods and cumulatively (Table 4). At 21 d, chickens fed YCW-, MP-, and BG-purified fractions increased (P ≤ 0.01) the villi height of jejunal mucosa with respect to chickens fed the control diets (Table 5). The intestine and pancreas RW at 21 d were not affected by treatment. However, chickens fed YCW had lower (P ≤ 0.01) liver RW than those fed the
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Ingredient Wheat Barley Rye Soybean meal (48% CP) Full-fat soybean Animal fat Calcium carbonate Dicalcium phosphate dl-Methionine (99.0%) l-Lysine·HCl (78.8%) l-Threonine (98.5%) NaCl Choline (50%) Minerals and vitamins1 Yeast fraction or carried2 Total Calculated analysis ME (kcal/kg) CP (%) Total Lys (%) Total Met (%) Total Met + Cys (%) Total Thr (%) Ca (%) Total P (%) Available P (%) Analyzed nutrients CP (%) Crude fat (%) DM (%)
Experiment 2
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Table 2. Effect of AVI, YCW, MP, and BG adding to broiler diets on growth performances1 (experiment 1) Experimental diets2 Item
677 2,244
AVI 668 2,361
YCW
MP
651 2,313
BG
655 2,284
661 2,312
MP + BG
SEM
Probability
647 2,310
9.97 37.2
0.22 0.43
30.1 74.6 52.3
29.7 80.6 55.1
28.8 79.2 54.0
29.0 77.6 53.3
29.4 78.6 53.9
28.6 79.0 53.9
0.47 1.64 0.88
0.22 0.35 0.43
44.0 134.2 87.7
43.2 139.8 90.1
43.8 136.8 89.4
42.8 135.1 87.7
42.6 138.5 89.0
43.1 137.3 89.1
0.80 3.49 1.82
0.85 0.79 0.83
1.464c 1.800 1.675
1.458c 1.733 1.633
1.520a 1.726 1.654
1.474bc 1.741 1.645
1.452c 1.762 1.650
1.507ab 1.735 1.652
0.013 0.022 0.014
0.01 0.26 0.70
1.4
3.7
5.8
4.3
1.4
6.5
1.38
0.38
a–c
Means within a row lacking common superscripts differ significantly (P ≤ 0.05). DWG = daily weight gain; DFI = daily feed intake; FCR = feed conversion ratio; n = 6 replicates of 23 chickens each from 0 to 21 d and 13 chickens each from 22 to 42 d. 2 AVI = avilamycin at 10 mg/kg of feed; YCW = yeast cell walls at 500 mg/kg of feed; MP = mannoprotein at 95 mg/kg of feed; BG = β-glucans at 145 mg of BG/kg of feed. 1
control and MP diets but no differences were observed with respect to chickens fed with BG (Table 5).
DISCUSSION Data obtained in these studies did not show significant positive effects of AVI, YCW, and its components on growth performance parameters of broiler chickens. We hypothesized that the lack of significant effect of AVI on growth performance parameters might be due to the absence of a real microbial challenge. It is well know that the lack of a negative environment that does not represent any challenge to the host clearly limits the response of dietary AGP (Bedford, 2000). Contrary to our results, dietary YCW have been shown to improve broiler chicken productivity (Santin et al., 2001; Zhang et al., 2005; Baurhoo et al., 2007). The MOS are indigestible to nonruminant animals and can provide competitive binding sites for pathogenic digestive bacteria, decreasing their intestinal attachment and colonization (Newman, 1994). The BG has been shown
to be protective in several disease challenge studies in animals (Mansell et al., 1978; Williams and Di Luzio, 1979; Reynolds et al., 1980). Apparently, the use of MOS, BG, or YCW can bring more benefit in animals maintained under microbial challenge conditions. In broilers reared in cages, Huff et al. (2006) observed an improvement on BW and FCR of broiler fed 20 m of BG/kg of feed and challenged with Escherichia coli. On the contrary, nonchallenged chickens fed BG reduced the BW (Huff et al., 2006). In another study (Chae et al., 2006), the positive effects of BG on chicken productivity were more consistent in birds maintained in litter pens than those reared in cages (Chae et al., 2006). Experiments without challenge conditions showed improvements in productivity of broilers maintained in cages and fed 50 mg of BG/kg of feed (Zhang et al., 2008). The structure, molecular weight, and origin are also factors related with the effectiveness of the BG in poultry diets. Regarding MP, there is a lack of information about the MP use in broiler feeding. Oyofo et al. (1989a,b,c) performed some studies in broilers chickens
Table 3. Effect of dietary adding of AVI, YCW, MP, and BG fractions on lymphoid organ relative weights1 of broiler chickens (experiment 1) Experimental diets2 Item Relative weight (% of BW) Spleen Bursa of Fabricius Thymus a–c
Control 0.115 0.216 0.452bc
AVI
YCW
0.118 0.262 0.433c
0.138 0.248 0.567ab
MP 0.123 0.265 0.459bc
BG 0.143 0.271 0.489abc
MP + BG
SEM
Probability
0.149 0.266 0.582a
0.011 0.021 0.037
0.18 0.46 0.05
Means within a row lacking common superscripts differ significantly (P ≤ 0.05). n = 8 organs. 2 AVI = avilamycin at 10 mg/kg of feed; YCW = yeast cell walls at 500 mg/kg of feed; MP = mannoprotein at 95 mg/kg of feed; BG = β-glucans at 145 mg of BG/kg of feed. 1
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BW (g/bird) d 21 d 42 DWG (g/d) 0 to 21 d 22 to 42 d 0 to 42 d DFI (g/d) 0 to 21 d 22 to 42 d 0 to 42 d FCR (g/g) 0 to 21 d 22 to 42 d 0 to 42 d Mortality (%) 0 to 42 d
Control
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Table 4. Effect of YCW, MP, and BG adding to broiler diets on growth performances (experiment 2) Experimental diets2 Item
717 2,404
YCW
MP
723 2,431
BG
748 2,419
SEM
733 2,430
14.2 40.5
Probability 0.70 0.90
32.0 80.3 56.8
32.3 81.3 56.8
33.5 79.6 56.5
32.8 80.8 56.2
0.67 1.80 0.86
0.69 0.90 0.90
52.3 134.1 93.1
51.9 131.6 91.8
52.6 128.4 90.4
51.2 128.4 89.8
1.18 2.15 1.4
0.85 0.23 0.26
0.029 0.027 0.024
0.30 0.29 0.20
1.96
0.18
1.635 1.672 1.660
1.608 1.619 1.615
13.2
1.569 1.615 1.600
9.3
1.562 1.595 1.584
12.7
13.2
1
DWG = daily weight gain; DFI = daily feed intake; FCR = feed conversion ratio; n = 6 replicates of 40 chickens each from 0 to 21 d and 33 chickens each from 22 to 42 d. 2 YCW = yeast cell walls at 500 mg/kg of feed; MP = mannoprotein at 190 mg/kg of feed; BG = β-glucans at 227 mg/kg of feed.
and evaluated the efficacy of purified fractions of mannose at 2% or 20 g/kg of feed to control Salmonella Typhimurium colonization rather than on increasing chicken productivity. In our studies, the MP (95 and 190 mg of MP/kg of feed in experiment 1 or 2, respectively) fractions were included at very low dosages in diets of broiler chickens that were not challenged. In experiment 1, chickens fed MP + BG had similar RW of thymus as chickens fed YCW but greater than those fed control diet with and without AVI. According to Ferket et al. (2002), the YCW components might stimulate the gut-associated immune system by acting as a nonpathogenic microbial antigen, giving an adjuvant-like effect. Studies in mammals have described the importance of digestive microbial antigen stimulation on the development of lymphoid organ tissue (Bealmear, 1980; Pabst et al., 1998; Rothkötter et al., 1991). In contrast, AGP avoid bacterial overgrowing and reduce the bacterial immune stimulation
at an intestine level. In our studies, YCW and MP + BG induced some effects on lymphoid organ tissues of chickens. Studies of dietary YCW and MP on relative lymphoid organ weights of broiler are limited or missing. Ao et al. (2004) did not find any effect on bursa of Fabricius RW of broilers fed 1.5 g of YCW (MOS)/ kg of feed. Guo et al. (2003) and Zhang et al. (2008) observed an increase of spleen, bursa of Fabricius, and thymus RW of broilers fed diets supplemented with 40 or 50 mg of BG/kg of feed. In agreement with our results, Shafey et al. (2001) and Santin et al. (2003) did not observe any differences in antibody response of NDV vaccine in broiler chickens receiving YCW or YCW identified as MOS. However, broilers fed YCW such as MOS and BG improved the antibody response of infectious bursal disease virus vaccine (Shashidhara and Devegowda, 2003) and red blood cells (Guo et al., 2003), respectively. In experiment 2, YCW, MP, and BG significantly increased the villus height of jejunal
Table 5. Effect of YCW, MP, and BG adding to broiler diets on mucosa villus height (jejunum)1 and digestive organ relative weights2 (experiment 2) Experimental diets3 Item Villus height (µm) Relative weight1 (% of BW) Intestine Liver Pancreas a,b
Control b
957
9.5 3.4a 0.412
YCW 1,159
a
8.9 2.9b 0.368
MP
BG
a
a
1,156
9.4 3.4a 0.398
1,090
9.7 3.1ab 0.381
SEM 39.1 0.25 0.01 0.018
Probability 0.01 0.09 0.01 0.21
Means within a row lacking common superscripts differ significantly (P ≤ 0.05). n = 10 jejunum samples. 2 n = 10 organs. 3 YCW = yeast cell walls at 500 mg/kg of feed; MP = mannoprotein at 190 mg/kg of feed; BG = β-glucans at 227 mg/kg of feed. 1
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BW (g/bird) d 21 d 42 DWG (g/d) 0 to 21 d 22 to 42 d 0 to 42 d DFI (g/d) 0 to 21 d 22 to 42 d 0 to 42 d FCR (g/g) 0 to 21 d 22 to 42 d 0 to 42 d Mortality (%) 0 to 42 d
Control
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ACKNOWLEDGMENTS We thank Nicolas Andrea, Faculty of Veterinary Medicine, Ghent University, Ghent, Belgium, for cooperation in the histological study of the present paper. The support of Comissió Interdepartamental per a la Recerca i Tecnologia (CIRIT, 2005 SGR-00720) is also acknowledged.
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Baurhoo, B., L. Phillip, and C. A. Ruiz-Feria. 2007. Effects of purified lignin and mannan oligosaccharides on intestinal integrity and microbial populations in ceca and litter of broiler chickens. Poult. Sci. 86:1070–1078. Bealmear, P. M. 1980. Host defence mechanisms in gnotobiotic animal. Pages 261–350 in Immunologic Defects in Laboratory Animals. Vol. 2. E. Gershwin and B. Marchant, ed. Plenum Press, New York, NY. Bedford, M. R. 2000. Removal of antibiotic growth promoters from poultry diets: Implications and strategies to minimise subsequent problems. World’s Poult. Sci. J. 56:347–365. Chae, B. J., J. D. Lohakare, W. K. Moon, S. L. Lee, Y. H. Park, and T.-W. Hahn. 2006. Effects of supplementation of β-glucan on the growth performance and immunity in broilers. Res. Vet. Sci. 80:291–298. DIN. 1977. DIN 51900. Determination of the gross calorific value by the bomb calorimeter and calculation of the net calorific value. Deutsches Institut für Normung e.V., Berlin, Germany. EURASYP. 2007. Yeast products: Yeast cell wall. http://www.eurasyp.org/public.levure.ecorce.screen#composants Accessed July 2008. Ferket, P. R., C. W. Parks, and J. L. Grimes. 2002. Benefits of dietary antibiotic and mannanoligosaccharide supplementation for poultry. In Proc. Multi-State Poultry Feeding and Nutrition Conference, Indianapolis, IN. http://ag.ansc.purdue.edu/poultry/multistate/Proceedings.Multi-State%20MeetingFerket.pdf Accessed June 2006. Guo, Y., R. A. Ali, and A. M. Qureshi. 2003. The influence of β-glucan on immune response in broiler chicks. Immunopharmacol. Immunotoxicol. 25:461–472. Hooge, D. M. 2004. Meta-analysis of broiler chicken pen trials evaluating dietary mannan oligosaccharide, 1993–2003. Int. J. Poult. Sci. 3:163–174. Huff, G. R., W. E. Huff, N. C. Rath, and G. Tellez. 2006. Limited treatment with β-1, 3/1, 6-glucan improves production values of broiler chickens challenged with Escherichia coli. Poult. Sci. 85:613–618. Mansell, P. W. A., G. Rowden, and C. Hammer. 1978. Clinical experiences with the use of glucan. Pages 255–280 in Immune Modulation and Control of Neoplasia by Adjuvant Therapy. Raven Press, New York, NY. NRC. 1994. Nutrient Requirements of Poultry. National Academy Press, Washington, DC. Newman, K. 1994. Mannan-oligosaccharides: Natural polymers with significant impact on the gastrointestinal microflora and the immune system. Pages 167–174 in Biotechnology in the Feed Industry: Proceedings of Alltech’s Tenth Annual Symposium. T. P. Lyons and K. A. Jacques, ed. Nottingham University Press, Nottingham, UK. Oyofo, B. A., J. R. DeLoach, D. E. Corrier, J. O. Norman, R. L. Ziprin, and H. H. Mollenhauer. 1989a. Effect of carbohydrates on Salmonella Typhimurium colonization in broiler chickens. Avian Dis. 33:531–534. Oyofo, B. A., J. R. DeLoach, D. E. Corrier, J. O. Norman, R. L. Ziprin, and H. H. Mollenhauer. 1989b. Prevention of Salmonella Typhimurium colonization of broilers with d-mannose. Poult. Sci. 68:1357–1360. Oyofo, B. A., R. E. Droleskey, J. O. Norman, H. M. Mollenhauer, R. L. Ziprin, D. E. Corrier, and J. R. DeLoach. 1989c. Inhibition by mannose of in vitro colonization of chicken small intestine by Salmonella Typhimurium. Poult. Sci. 68:1351–1356. Pabst, R., M. Geist, H. J. Rothkötter, and F. J. Fritz. 1998. Postnatal development and lymphocyte production of jejunal and ileal Peyer’s patches in normal and gnotobiotic pigs. Immunology 64:539–544. Raa, J. 2003. The use of immune-stimulant to enhance disease resistance and growth performance of fish and shrimp. Pages 67–75 in Proc. XI Congreso Nacional de AMENA y I Congreso Latino-Americano de Nutrición Animal, Cancún, Qroo (México). Asociación Mexicana de Especialistas en Nutrición Animal, A.C. Querétaro, Qro. México. Reynolds, J. A., M. D. Kastello, D. G. Harrington, C. L. Crabbs, C. J. Peters, J. V. Jemski, G. H. Scott, and N. R. DiLuzio. 1980.
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mucosa of chickens. These results not only were in agreement with other studies in broilers (Santin et al., 2001; Zhang et al., 2005; Baurhoo et al., 2007) but also showed that dietary MP and BG increased mucosal villus height. We do not have a clear explanation regarding the effect of lighter liver RW observed in chickens fed YCW and BG (experiment 2). The dietary YCW and BG could represent an indirect benefit in terms of minor liver damage. In fact, the greater but not significant broiler mortality observed in experiment 2 was due to heat stress problems that occurred at 36 d of age. In broilers, the increment of liver weight can be reflective of an acute phase of an inflammatory response because the liver is the site for the synthesis of acute phase proteins (Xie et al., 2000). Huff et al. (2006) reported lighter liver RW in broilers challenged with E. coli and fed BG in the diet. In experiment 1, the effect of YCW and MP + BG of reducing the feed efficiency of chickens could be a consequence of their effect of promoting mucosal growth and immune stimulation and the use of a single diet low in some essential amino acids. Our data did not show clear improvements of YCW, BG, MP, and MP + BG on animal growth performance and other variables in broiler chickens. However, we can suggest that the effect on thymus and liver relative weights and villus morphology of broiler chickens were induced with the same intensity by the use of complete YCW, MP + BG, and BG. The MP + BG and YCW increased the relative thymus weights of chickens. The YCW and BG maintained lighter relative liver weight in chickens. The YCW increased villus height of jejunal mucosa, and this effect was also obtained with the single use of MP- and BG-purified fractions. More research is needed using YCW and purified MP and BG to further clarify their possible beneficial effects on broiler performances.
YEAST CELL WALLS; β-1, 3/1, 6-GLUCANS; AND MANNOPROTEINS
Spring, P., C. Wenk, A. K. Dawson, and E. K. Newman. 2000. The effects of dietary mannanoligosaccharides on cecal parameters and the concentrations of enteric bacteria in the ceca of Salmonella-challenged broilers chicks. Poult. Sci. 79:205–211. Takahashi, K., S. Konashi, Y. Akiba, and M. Hirioguchi. 1994. Effects of dietary threonine level on antibody production. Anim. Sci. Technol. (Jpn.) 65:956–960. Williams, D. L., and N. R. DiLuzio. 1979. Glucan induced modification of experimental Staphylococcus aureus infection in normal, leukemic and immunosuppressed mice. Adv. Exp. Med. Biol. 121:291–306. Xie, H., N. C. Rath, G. R. Huff, W. E. Huff, and J. M. Balog. 2000. Effects of Salmonella Typhimurium lipopolysaccharide on broiler chickens. Poult. Sci. 70:33–40. Zhang, B., Y. Guo, and Z. Wang. 2008. The modulation effect of β-1, 3/1, 6-glucan supplementation in the diet on performance and immunological response of broiler chickens. Asian-austalas. J. Anim. Sci. 21:237–244. Zhang, A. W., B. D. Lee, S. K. Lee, K. W. Lee, G. H. An, K. B. Song, and C. H. Lee. 2005. Effects of yeast (Saccharomyces cerevisiae) cell components on growth performance, meat quality, and ileal mucosa development of broilers chicks. Poult. Sci. 84:1015–1021.
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Glucan-induced enhancement of host resistance to selected infectious diseases. Infect. Immun. 30:51–57. Rosen, G. D. 2007. Halo-analysis of the efficacy of Biol.-Mos® in broiler nutrition. Br. Poult. Sci. 48:21–26. Rothkötter, H. J., H. Ulrich, and R. Pabst. 1991. The postnatal development of gut lamina propia lymphocytes: Number, proliferation, and T and B cell subsets in conventional and germ-free pigs. Pediatr. Res. 29:237–242. Santin, E., A. Maiorka, and M. Macari. 2001. Performance and intestinal mucosa development of broilers chickens fed diets containing Saccharomyces cerevisiaes cell wall. J. Appl. Poult. Res. 10:236–244. Santin, E., A. C. Paulillo, A. Majorka, L. S. N. Okada, M. Macari, A. V. Fischer da Silva, and A. C. Alessi. 2003. Evaluation of the efficacy of Saccharomyces cerevisiae cell wall to ameliorate the toxic effects of aflatoxin in broilers. Int. J. Poult. Sci. 5:341–344. SAS Institute. 1996. SAS User’s Guide. Relase 6.12 ed. SAS Institute Inc., Cary, NC. Shafey, T. M., S. Al-Mufarej, G. B. Havenstein, and A. J. Jarelnabi. 2001. Effects of mannan oligosaccharides on antibody response to infectious bronchitis, infectious bursal disease and Newcastle disease in chickens. J. Appl. Anim. Res. 19:117–127. Shashidhara, R. G., and G. Devegowda. 2003. Effect of dietary mannan oligosaccharide on broiler breeders production traits and immunity. Poult. Sci. 82:1319–1325.
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Key words: mannoprotein complex, β-1, 3/1, 6-glucan, yeast cell wall, broiler chicken, villi height 2009 Poultry Science 88:601–607 doi:10.3382/ps.2008-00298 of mannan sugar polymers), 15 to 30% proteins, 5 to 20% lipids, and no more than 5% of chitin (AguilarUscanga and François, 2003; EURASYP, 2007). Most of the protein is linked to the mannanoligosaccharides (MOS) and is referred to as the mannoprotein complex (MP). In the digestive tract of animals, MOS present in YCW could act as high-affinity ligands, with the potential benefit of offering a competitive binding site for pathogenic bacteria mannose-specific type-1 fimbriae (Spring et al., 2000). The β-1, 3/1, 6-glucans also present in YCW should be recognized as an immune modulator substance in animals and humans (Abel and Czop, 1992; Raa, 2003); thus, dietary YCW might exert some benefits on the immune system of intestinal mucosa (Ferket et al., 2002). The importance of β-1, 3/1, 6-glucans and MP-purified fractions on the mode of action for YCW additives has not been studied in poultry. Thus, the aim of these studies was to evaluate the effect of YCW obtained from S. cerevisiae of
INTRODUCTION Saccharomyces cerevisiae yeast cell wall (YCW) components have been used in animal feeding since the last decades (Hooge, 2004; Rosen, 2007). Their inclusion in broiler diets has resulted in improvements of animal productivity, which was attributed to physiological effects on intestinal digestive mucosa (Santin et al., 2001; Zhang et al., 2005; Baurhoo et al., 2007). However, the mode of action of YCW products in broiler chicken diets is not well understood and the characteristics of YCW products have been poorly defined. Typically, commercial YCW are composed of 30 to 60% polysaccharides (15 to 30% of β-1, 3/1, 6-glucan and 15 to 30% ©2009 Poultry Science Association Inc. Received July 22, 2008. Accepted October 27, 2008. 1 Corresponding author: [email protected]
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BG treatments. In experiment 2, there was a group-fed basal diet and 3 additional experimental groups receiving the basal diet supplemented with, respectively: 500 mg of YCW/kg of feed, 190 mg of MP/kg of feed, and 227 mg of BG/kg of feed. At 42 d of age, no difference in broiler growth rates was observed. Samples of the jejunum were collected at 21 d of age to determine villus height. Significantly higher villus height was observed in YCW, MP, and BG groups compared with that of the control group. The relative percentage of liver weight (P ≤ 0.01) was lower in broilers fed YCW than in broilers fed the control diet, but no differences were observed in respect to chickens fed BG. Data of these studies suggested that the changes in thymus and liver relative weights and villus morphology of broilers were induced with the same intensity by the use of complete YCW, MP + BG, and BG supplements.
ABSTRACT Two experiments were carried out to evaluate the effect of dietary addition of yeast cell wall (YCW); β-1, 3/1, 6-glucan (BG); and mannoprotein complex (MP) purified fractions in broilers. In experiment 1, there was a control diet and 5 experimental diets containing, respectively: 10 mg of avilamycin (AVI)/ kg of feed, 500 mg of YCW/kg of feed, 95 mg of MP/ kg of feed, 145 mg of BG/kg of feed, and 95 mg of MP plus 145 mg of BG/kg of feed. All birds were vaccinated via drinking water against Newcastle disease virus at 9 d of age. At 42 d, chickens fed AVI, YCW, MP + BG, and BG diets had similar BW not significantly different from chickens fed the control diet. The antibody response of Newcastle disease virus vaccine was not affected by any experimental treatment. Broilers fed MP + BG diet had greater thymus weights (P ≤ 0.05), as a percentage of BW than those from the control and AVI treatments, but similar with respect to YCW and
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the yeast extract industry and their MP and β-1, 3/1, 6-glucans-purified fractions supplemented in the diets of broiler chickens.
MATERIALS AND METHODS Animal Housing
Diets The YCW efficacy was tested on wheat-based diets in mash form without any antibiotic growth promoters (AGP) and coccidiostats (Table 1). There were no exogenous enzymes used for wheat, barley, and rye cereals because they might mask the effects of yeast additives. Diets were formulated to meet or exceed NRC (1994) requirements. In experiment 1, a single diet was used throughout the 42 d of trial to avoid adaptation to the change of diet. In experiment 2, starter and grower diets were used from 0 to 21 and from 21 to 42 d, respectively (Table 1). The analytical composition of the main ingredients and feeds was determined using standard methods (AOAC, 1990) including DM (method 934.01), CP (method 968.06; protein-nitrogen determination, LECO FP-528, St. Joseph, MI), crude fat (method 920.39), and gross energy (DIN, 1977) using an adiabatic calorimeter bomb (Calorimeter C-4000 A, IKA Analysentrchnik GmbH, Heitersheim, Germany). In experiment 1, experimental YCW contained 19% mannose and 29% of β-1, 3/1, 6-glucans. In experiment 2, experimental YCW contained 21% of mannose and 25% of β-1, 3/1, 6-glucans; purified MP contained 55% mannose; and purified β-glucans (BG) contained 55% of β-1, 3/1, 6-glucans in β-1, 3 main forms. The determination of glucans and mannan polysaccharides of YCW, MP, and BG was conducted at Bio-Springer (Maisons-Alfort Cedex, France) by using the EURASYP (2007) methodology. In both experiments, chickens were weighed at 1, 21, and 42 d of age. The BW, daily weight gain, daily feed intake, and feed conversion ratio (FCR) were calculated for each period and for the overall experiment.
Experiment 1 evaluated the effect of YCW from the yeast extract industry (Lesaffre Feed Additives, Marquette-Lez-Lille, France) and their main polysaccharide components such as MP- and BG-purified fractions on growth performance parameters, antibody response against Newcastle disease virus (NDV) vaccine, and lymphoid organs (spleen, thymus, and bursa of Fabricius) relative weights (RW). A total of 828 broilers chickens were used. There was a control diet and 5 experimental diets containing, respectively: 10 mg of avilamycin (AVI)/kg of feed, 500 mg of YCW/kg of feed, 95 mg of MP/kg of feed, 145 mg of BG/kg of feed, and 95 mg of MP plus 145 mg of BG/kg of feed. Each treatment was replicated 6 times of 23 birds. At 9 d of age, all birds were vaccinated through drinking water with an attenuated NDV vaccine (“La Sota” strain). At 23 and 36 d, 3 chickens per pen (or 18 per treatment) were bled through the brachial vein to measure antibody titters against NDV by the hemagglutination inhibition test (Takahashi et al., 1994). At 36 d of age, 8 chickens from each treatment were selected at random and were killed according to protocols accepted by the Ethical Commission of Institute of Research, Food Technology and Agriculture (Catalonia, Spain). The spleen, bursa of Fabricius, and thymus (all right and left lobules) were excised and weighed. Afterwards, adherent fat from these organs was removed and their RW expressed as a percentage of live BW were calculated: RW = organ weight × 100/chicken BW.
Experiment 2 Experiment 2 evaluated the effect of YCW from the yeast extract industry, MP, and BG on growth performance, villus height of jejunal mucosa, and RW of digestive organs (intestine, pancreas, and liver). A total of 800 broiler chicks were used, and there was a control diet and 3 experimental diets containing, respectively: 500 mg of YCW/kg of feed, 190 mg of MP/kg of feed, and 227 mg of BG/kg of feed. Each treatment included 5 replicates of 40 chickens. At 21 d of age, 10 chickens from each treatment were killed. The complete intestine (from the gizzard-duodenal junction to rectum-cloacal junction), pancreas, and liver were excised and weighed. Afterward, adherent fat from these organs was removed and their RW were calculated. From the same killed chicks, 10 samples from each experimental treatment were collected from the proximal jejunum (5 cm) 2 cm following the end of the duodenal loop. Samples from jejunum were fixed in Carnoy’s solution (60% absolute ethanol + 30% chlorophorm + 10% glacial acetic acid-based solution), dehydrated (70% alcohol, 60 min; 96% alcohol, 45 min and then 60 min; isopropylic alcohol, 60 min), embedded in paraffin, and 4-μm sections were cut. Then, sections
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Two experiments were conducted using commercial 1-d-old Ross 308 male broiler chicks. Chickens were placed in 36 one-square meter cages in the first experiment (experiment 1) and in 20 floor pens of 4 square meters with wood shavings as litter in the second experiment (experiment 2). There were 6 and 5 blocks in experiment 1 and 2, respectively, corresponding to location in the experimental house, and treatments were randomly distributed within each block. The experimental house was provided with forced ventilation, artificial light, and gas heating. The temperature inside the house on arrival was 33 to 35°C and was decreased by 3°C each week until 22°C. The lighting program was 23 h of light the first 4 d, 20 h until 10 d, and 18 afterwards. Feed and water were provided for ad libitum consumption throughout the 42 d of experimentation.
Experiment 1
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Table 1. Ingredient composition (g/kg) and calculated nutrient analysis of experimental diets Experiment 1 Item
Single diet
Starter diet
Grower diet
437.2 150.0 50.0 185.5 109.7 30.0 11.81 14.51 2.61 1.15 — 3.00 0.05 4.00 0.50 1,000.0
379.8 124.9 50.0 232.3 120.0 48.0 11.90 17.10 2.70 2.20 0.80 3.20 1.10 4.00 2.00 1,000.0
519.6 63.2 25.0 152.6 140.0 58.0 10.30 16.00 2.10 2.70 0.70 3.20 0.60 4.00 2.00 1,000.0
3,000 20.0 1.10 0.55 0.90 0.72 0.90 0.60 0.40
3,000 22.0 1.27 0.58 0.94 0.85 1.00 0.68 0.45
3,150 20.0 1.15 0.50 0.83 0.76 0.90 0.64 0.42
20.3 6.2 89.7
22.6 8.3 89.5
19.3 9.2 89.7
1 Provided per kilogram of feed: vitamin A, 12,000 IU; vitamin D3, 2,400 IU; vitamin E, 30 mg; vitamin K3, 3 mg; thiamine, 2.2 mg; riboflavin, 8 mg; pyridoxine, 5 mg; cyanocobalamin, 11 μg; folic acid, 1.5 mg; biotin, 150 μg; calcium pantothenate, 25 mg; nicotinic acid, 65 mg; Mn (manganese sulfate), 60 mg; Zn (zinc oxide), 40 mg; I (potassium iodate), 0.33 mg; Fe (ferrous sulfate), 80 mg; Cu (copper sulfate), 8 mg; Se (sodium selenite), 0.15 mg; ethoxyquin, 150 mg. 2 The experimental yeast fractions: 500 g of yeast cell walls (YCW), 95 g of mannoprotein (MP), and 145 g of β-glucans (BG) in experiment 1 and 500 g of YCW, 190 g of MP, and 227 g of BG in experiment 2 were included per ton of experimental feed to expenses of corn starch.
were stained with Luxol Fast Blue/PAS stain (Polysciences Inc., Ajalvir-Madrid, Spain) to determine the villus height in at least 3 villi per jejunal sample.
Statistical Analysis Experiment 1 was analyzed by 2-way ANOVA as a randomized complete block design with 6 blocks corresponding to location within the house and 6 dietary treatments; experiment 2 was analyzed by the same statistical design as experiment 1 but with 5 blocks and 4 dietary treatments (SAS Institute, 1996). Differences between treatments were established by Tukey’s test at P ≤ 0.05 probability. The data expressed as percentages such as mortality were transformed to arcsine values.
RESULTS Experiment 1 Experimental diets did not affect growth performance parameters at different periods and cumulatively ex-
cept for FCR from 0 to 21 d (Table 2). The FCR was impaired (P ≤ 0.01) in chickens fed YCW and MP + BG compared with those fed the control diet, AVI, and BG. At 38 d, chickens fed MP + BG fractions had significantly heavier thymus RW (P ≤ 0.05) with respect to those fed the control and the AVI diets, and similar than those chicks fed YCW and BG (Table 3), and the bursa of Fabricius and spleen RW were not affected by treatment (Table 3). At 23 and 36 d, the antibody response of NDV vaccine did not show changes due to the use of experimental diet.
Experiment 2 Experimental diets did not affect growth performance parameters at different periods and cumulatively (Table 4). At 21 d, chickens fed YCW-, MP-, and BG-purified fractions increased (P ≤ 0.01) the villi height of jejunal mucosa with respect to chickens fed the control diets (Table 5). The intestine and pancreas RW at 21 d were not affected by treatment. However, chickens fed YCW had lower (P ≤ 0.01) liver RW than those fed the
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Ingredient Wheat Barley Rye Soybean meal (48% CP) Full-fat soybean Animal fat Calcium carbonate Dicalcium phosphate dl-Methionine (99.0%) l-Lysine·HCl (78.8%) l-Threonine (98.5%) NaCl Choline (50%) Minerals and vitamins1 Yeast fraction or carried2 Total Calculated analysis ME (kcal/kg) CP (%) Total Lys (%) Total Met (%) Total Met + Cys (%) Total Thr (%) Ca (%) Total P (%) Available P (%) Analyzed nutrients CP (%) Crude fat (%) DM (%)
Experiment 2
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Table 2. Effect of AVI, YCW, MP, and BG adding to broiler diets on growth performances1 (experiment 1) Experimental diets2 Item
677 2,244
AVI 668 2,361
YCW
MP
651 2,313
BG
655 2,284
661 2,312
MP + BG
SEM
Probability
647 2,310
9.97 37.2
0.22 0.43
30.1 74.6 52.3
29.7 80.6 55.1
28.8 79.2 54.0
29.0 77.6 53.3
29.4 78.6 53.9
28.6 79.0 53.9
0.47 1.64 0.88
0.22 0.35 0.43
44.0 134.2 87.7
43.2 139.8 90.1
43.8 136.8 89.4
42.8 135.1 87.7
42.6 138.5 89.0
43.1 137.3 89.1
0.80 3.49 1.82
0.85 0.79 0.83
1.464c 1.800 1.675
1.458c 1.733 1.633
1.520a 1.726 1.654
1.474bc 1.741 1.645
1.452c 1.762 1.650
1.507ab 1.735 1.652
0.013 0.022 0.014
0.01 0.26 0.70
1.4
3.7
5.8
4.3
1.4
6.5
1.38
0.38
a–c
Means within a row lacking common superscripts differ significantly (P ≤ 0.05). DWG = daily weight gain; DFI = daily feed intake; FCR = feed conversion ratio; n = 6 replicates of 23 chickens each from 0 to 21 d and 13 chickens each from 22 to 42 d. 2 AVI = avilamycin at 10 mg/kg of feed; YCW = yeast cell walls at 500 mg/kg of feed; MP = mannoprotein at 95 mg/kg of feed; BG = β-glucans at 145 mg of BG/kg of feed. 1
control and MP diets but no differences were observed with respect to chickens fed with BG (Table 5).
DISCUSSION Data obtained in these studies did not show significant positive effects of AVI, YCW, and its components on growth performance parameters of broiler chickens. We hypothesized that the lack of significant effect of AVI on growth performance parameters might be due to the absence of a real microbial challenge. It is well know that the lack of a negative environment that does not represent any challenge to the host clearly limits the response of dietary AGP (Bedford, 2000). Contrary to our results, dietary YCW have been shown to improve broiler chicken productivity (Santin et al., 2001; Zhang et al., 2005; Baurhoo et al., 2007). The MOS are indigestible to nonruminant animals and can provide competitive binding sites for pathogenic digestive bacteria, decreasing their intestinal attachment and colonization (Newman, 1994). The BG has been shown
to be protective in several disease challenge studies in animals (Mansell et al., 1978; Williams and Di Luzio, 1979; Reynolds et al., 1980). Apparently, the use of MOS, BG, or YCW can bring more benefit in animals maintained under microbial challenge conditions. In broilers reared in cages, Huff et al. (2006) observed an improvement on BW and FCR of broiler fed 20 m of BG/kg of feed and challenged with Escherichia coli. On the contrary, nonchallenged chickens fed BG reduced the BW (Huff et al., 2006). In another study (Chae et al., 2006), the positive effects of BG on chicken productivity were more consistent in birds maintained in litter pens than those reared in cages (Chae et al., 2006). Experiments without challenge conditions showed improvements in productivity of broilers maintained in cages and fed 50 mg of BG/kg of feed (Zhang et al., 2008). The structure, molecular weight, and origin are also factors related with the effectiveness of the BG in poultry diets. Regarding MP, there is a lack of information about the MP use in broiler feeding. Oyofo et al. (1989a,b,c) performed some studies in broilers chickens
Table 3. Effect of dietary adding of AVI, YCW, MP, and BG fractions on lymphoid organ relative weights1 of broiler chickens (experiment 1) Experimental diets2 Item Relative weight (% of BW) Spleen Bursa of Fabricius Thymus a–c
Control 0.115 0.216 0.452bc
AVI
YCW
0.118 0.262 0.433c
0.138 0.248 0.567ab
MP 0.123 0.265 0.459bc
BG 0.143 0.271 0.489abc
MP + BG
SEM
Probability
0.149 0.266 0.582a
0.011 0.021 0.037
0.18 0.46 0.05
Means within a row lacking common superscripts differ significantly (P ≤ 0.05). n = 8 organs. 2 AVI = avilamycin at 10 mg/kg of feed; YCW = yeast cell walls at 500 mg/kg of feed; MP = mannoprotein at 95 mg/kg of feed; BG = β-glucans at 145 mg of BG/kg of feed. 1
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BW (g/bird) d 21 d 42 DWG (g/d) 0 to 21 d 22 to 42 d 0 to 42 d DFI (g/d) 0 to 21 d 22 to 42 d 0 to 42 d FCR (g/g) 0 to 21 d 22 to 42 d 0 to 42 d Mortality (%) 0 to 42 d
Control
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Table 4. Effect of YCW, MP, and BG adding to broiler diets on growth performances (experiment 2) Experimental diets2 Item
717 2,404
YCW
MP
723 2,431
BG
748 2,419
SEM
733 2,430
14.2 40.5
Probability 0.70 0.90
32.0 80.3 56.8
32.3 81.3 56.8
33.5 79.6 56.5
32.8 80.8 56.2
0.67 1.80 0.86
0.69 0.90 0.90
52.3 134.1 93.1
51.9 131.6 91.8
52.6 128.4 90.4
51.2 128.4 89.8
1.18 2.15 1.4
0.85 0.23 0.26
0.029 0.027 0.024
0.30 0.29 0.20
1.96
0.18
1.635 1.672 1.660
1.608 1.619 1.615
13.2
1.569 1.615 1.600
9.3
1.562 1.595 1.584
12.7
13.2
1
DWG = daily weight gain; DFI = daily feed intake; FCR = feed conversion ratio; n = 6 replicates of 40 chickens each from 0 to 21 d and 33 chickens each from 22 to 42 d. 2 YCW = yeast cell walls at 500 mg/kg of feed; MP = mannoprotein at 190 mg/kg of feed; BG = β-glucans at 227 mg/kg of feed.
and evaluated the efficacy of purified fractions of mannose at 2% or 20 g/kg of feed to control Salmonella Typhimurium colonization rather than on increasing chicken productivity. In our studies, the MP (95 and 190 mg of MP/kg of feed in experiment 1 or 2, respectively) fractions were included at very low dosages in diets of broiler chickens that were not challenged. In experiment 1, chickens fed MP + BG had similar RW of thymus as chickens fed YCW but greater than those fed control diet with and without AVI. According to Ferket et al. (2002), the YCW components might stimulate the gut-associated immune system by acting as a nonpathogenic microbial antigen, giving an adjuvant-like effect. Studies in mammals have described the importance of digestive microbial antigen stimulation on the development of lymphoid organ tissue (Bealmear, 1980; Pabst et al., 1998; Rothkötter et al., 1991). In contrast, AGP avoid bacterial overgrowing and reduce the bacterial immune stimulation
at an intestine level. In our studies, YCW and MP + BG induced some effects on lymphoid organ tissues of chickens. Studies of dietary YCW and MP on relative lymphoid organ weights of broiler are limited or missing. Ao et al. (2004) did not find any effect on bursa of Fabricius RW of broilers fed 1.5 g of YCW (MOS)/ kg of feed. Guo et al. (2003) and Zhang et al. (2008) observed an increase of spleen, bursa of Fabricius, and thymus RW of broilers fed diets supplemented with 40 or 50 mg of BG/kg of feed. In agreement with our results, Shafey et al. (2001) and Santin et al. (2003) did not observe any differences in antibody response of NDV vaccine in broiler chickens receiving YCW or YCW identified as MOS. However, broilers fed YCW such as MOS and BG improved the antibody response of infectious bursal disease virus vaccine (Shashidhara and Devegowda, 2003) and red blood cells (Guo et al., 2003), respectively. In experiment 2, YCW, MP, and BG significantly increased the villus height of jejunal
Table 5. Effect of YCW, MP, and BG adding to broiler diets on mucosa villus height (jejunum)1 and digestive organ relative weights2 (experiment 2) Experimental diets3 Item Villus height (µm) Relative weight1 (% of BW) Intestine Liver Pancreas a,b
Control b
957
9.5 3.4a 0.412
YCW 1,159
a
8.9 2.9b 0.368
MP
BG
a
a
1,156
9.4 3.4a 0.398
1,090
9.7 3.1ab 0.381
SEM 39.1 0.25 0.01 0.018
Probability 0.01 0.09 0.01 0.21
Means within a row lacking common superscripts differ significantly (P ≤ 0.05). n = 10 jejunum samples. 2 n = 10 organs. 3 YCW = yeast cell walls at 500 mg/kg of feed; MP = mannoprotein at 190 mg/kg of feed; BG = β-glucans at 227 mg/kg of feed. 1
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BW (g/bird) d 21 d 42 DWG (g/d) 0 to 21 d 22 to 42 d 0 to 42 d DFI (g/d) 0 to 21 d 22 to 42 d 0 to 42 d FCR (g/g) 0 to 21 d 22 to 42 d 0 to 42 d Mortality (%) 0 to 42 d
Control
606
Morales-López et al.
ACKNOWLEDGMENTS We thank Nicolas Andrea, Faculty of Veterinary Medicine, Ghent University, Ghent, Belgium, for cooperation in the histological study of the present paper. The support of Comissió Interdepartamental per a la Recerca i Tecnologia (CIRIT, 2005 SGR-00720) is also acknowledged.
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