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Poult Sci 2006. 85:2176-2179
© 2006 Poultry Science Association
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METABOLISM AND NUTRITION

Effect of ß-Mannanase (Hemicell) on Growth Performance and Immunity of Broilers

X. T. Zou1, X. J. Qiao and Z. R. Xu

Animal Science College, Zhejiang University, HangZhou, 310029, P. R. China

1 Corresponding author: xtzou{at}zju.edu.cn


   ABSTRACT
TOP
 ABSTRACT
INTRODUCTION
 MATERIALS AND METHODS
 RESULTS
 DISCUSSION
 REFERENCES
 
Two hundred four broilers (1-d-old) were randomly allocated to 4 treatments, each of which had 3 pens of 17 chicks per pen and were used to investigate the effects of ß-mannanase (Hemicell) on growth performance and immunity. The chicks received the same basal diet based on corn-soybean meal and Hemicell was added to the basal diet at 0, 0.025, 0.05, and 0.075%, respectively. Weight of each replicate was determined at wk 0, 3, and 6 of age. There were no significant differences in average feed intake in the 0- to 3-wk and 0- to 6-wk periods, and no differences in serum IgA, or IgG concentrations. However, the addition of Hemicell significantly increased (P < 0.05) weight gain in the 4- to 6-wk and 0- to 6-wk periods. Feed conversion for the 0.025 and 0.05% groups was significantly greater (P < 0.05) than for the control group in the 4- to 6-wk and 0- to 6-wk periods. Hemicell significantly increased (P < 0.05) the serum IgM concentration in 3- and 6-wk-old broilers. Proliferation of T lymphocytes in 6-wk-old broilers for the 0.05% group was also improved (P < 0.05) significantly. The results indicate that Hemicell may improve growth performance and immunity of broilers.

Key Words: Hemicell • growth performance • immunity • broiler


   INTRODUCTION
TOP
 ABSTRACT
 INTRODUCTION
MATERIALS AND METHODS
 RESULTS
 DISCUSSION
 REFERENCES
 
The identification and alleviation of factors that inhibit nutrient utilization are necessary for successful poultry production. Among potential factors reducing nutrient bio-availability are the nonstarch polysaccharides in ingredients such as guar, soybean meal, and sesame meal. It occurs in the forms of glucomannans and galactomannans in plant cell walls. Patel and McGinnis (1985) found that ß-mannan significantly decreased egg production, egg weight, and feed intake in laying hens. There are also some experiments that demonstrated ß-mannan reduced insulin secretion (Sambrook and Rainbird, 1985) and glucose absorption in swine (Rainbird et al., 1984).

Hemicell is a fermentation product of Bacillus lentus; its active ingredient is ß-mannanase, which hydrolyzes ß-mannan. Daskiran et al. (2004) demonstrated that ß-mannanase improved feed:gain ratio and reduced water:feed ratio and dry fecal output of broilers by degrading ß-mannans. Jackson et al. (2004) reported that ß-mannanase inclusion at 80 million units per ton improved broiler gains and feed conversion. Broiler experiments using corn-soybean meal–based diets have demonstrated that ß-mannanase improved ME, growth, and feed efficiency in broilers by about 3% (McNaughton et al., 1998). Petty et al. (2002) reported that addition of 0.05% ß-mannanase increased average daily gain compared with that of pigs fed the control (no ß-mannanase supplementation) diets. Corn-soybean meal–based diets are the most popular for broilers in China. Because soybean meal contains ß-mannans such as ß-galactomannan and ß-glucomannan, addition of ß-mannanase may improve utilization of soybean meal. Lee et al. (2003) reported that the marked improvement in feed:gain ratio caused by ß-mannanase is presumably due to degradation of residual gum leading to reduced viscosity. Wu et al. (2005) reported that addition of ß-mannanase improved feed conversion by approximately 4.2% in hens fed a low-energy diet supplemented with 0.05% Hemicell.

Little research has been conducted to investigate the effect of Hemicell on immunity of broilers fed a corn-soy diet. The goal of this study was to evaluate the effect of Hemicell on feed intake, weight gain, feed conversion, relative immune organ weight, serum immunoglobulin concentration, and T-lymphocyte proliferation.


   MATERIALS AND METHODS
TOP
 ABSTRACT
 INTRODUCTION
 MATERIALS AND METHODS
RESULTS
 DISCUSSION
 REFERENCES
 
Experimental Birds and Dietary Composition
All procedures were approved by the Institutional Animal Care and Use Committee of Zhejiang University. Two hundred four 1-d-old Avine broilers (Beijing Dafa Chiatai Co. Ltd., China) were randomly allocated to 4 treatments, each of which had 3 pens of 17 chicks per pen. The temperature was maintained at 34 ± 1°C up to 7 d of age and then gradually decreased to 26 ± 1°C by 21 d of age, after which the chicks were maintained at room temperature. Lighting was continuous and water and feed were available ad libitum. The chicks received the same basal diet based on corn-soybean meal and Hemicell was added to the basal diet at 0, 0.025, 0.05, and 0.075%, respectively. The Hemicell was provided by ChemGen Co., Ltd. (Shanghai, China) and the activity of ß-mannanase was greater than 165 x 106 U/kg. Starter and grower diets were offered to the birds from 0 to 3 wk and from 4 to 6 wk of age, respectively. Nutrient levels of the diets (Table 1Go) were based on the NRC (1994). Feeds were analyzed for CP, calcium, and total phosphorus according to the methods of AOAC (1990).


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  		Table 1. Ingredients and nutrient composition of diets
 
Sampling, Testing, and Observations
Feed consumption was recorded weekly. Weight of each replicate was determined at 0, 3 and 6 wk of age. Feed conversion ratios were determined for each feeding period.

Two birds of each replicate were randomly selected and slaughtered at 3 and 6 wk of age. The thymus, spleen, and bursa of each bird were collected and weighed. Relative immune organs were calculated according to the following equation: relative immune organ weight = immune organ weight/BW.

Blood samples were obtained via vena cava puncture and fresh blood was poured into a vessel. After being separated naturally, the serum was poured into an Eppendorf centrifuge tube (10 mL) and centrifuged for 10 min (5,500 x g). Pure serum samples were aspirated by pipette and stored in 1.5-mL Eppendorf tubes at –70°C until analysis. Fresh blood from 1 bird in each replicate in the 0.05% Hemicell and control groups at 6 wk of age were collected for the lymphocyte and blastogenesis assay. Concentrations of serum IgA, IgG, and IgM were measured using a biochemistry autoanalyzer (RX Daytona, Shining Sun Technology, Beijing, China).

In vitro cellular immune response was measured using a lymphocyte and blastogenesis assay. Lymphocytes (T cells) from whole blood were tested for blastogenic response to concanavalin A (ConA, Sigma Chemical Co., St. Louis, MO) and lipopolysaccharide (LPS, from Escherichia coli, Sigma Chemical Co.) according to the method of Bendich et al. (1984). The lymphocytes were collected by density-gradient centrifugation at 686 x g for 30 min and washed 3 times with Hanks’ balanced salt solution. The lymphocytes were then resuspended in 10 mL of RPMI 1640 complete culture medium (Sangon Biological Engineering Technology and Services Co., Shanghai, China) supplemented with 10% (vol/vol) of heat-inactivated fetal calf serum, 100 U of penicillin/mL, 100 µg of streptomycin/mL, and 25 mM of N-(2-hydroxyethyl)-piperazine-N-2-ethane-sulfonic acid. The cells were detected by trypan blue dye exclusion and counted to adjust the density to 2 x 106 cells/mL of culture medium. Then, 100 µL of cell suspension and ConA (or LPS) were added to a 96-well microtiter plate to provide a final concentration of 5 µg of ConA (or LPS)/mL. No cell suspension was added to control wells. After cultures were incubated for 48 h in a incubator at 37°C and 5% CO2, 10 µL of 3-(4, 5- dimethylthiazolyl), 2, 5-diphenyl tetrazolium bromide (5 mg/mL, Sigma Chemical Co.) was added to each well, and the plates incubated for 4 h. One hundred microliters of dimethyl sulfoxide (0.04 mol/L) was added to each well to stop the reaction. The plates were placed at room temperature for 20 min and the results were reported as optical density at 570 nm by using an EIA Reader (DG-3022A, Bio-Rad, Hercules, CA).

Statistical Analysis
Data were subjected to 1-way ANOVA by using the GLM procedure in SPSS 11.5 for Windows (SPSS Inc., Chicago, IL). If differences in treatment means were detected by ANOVA, Duncan’s multiple range test was applied to separate means. Statements of statistical significance are based on a probability of (P < 0.05).


   RESULTS
TOP
 ABSTRACT
 INTRODUCTION
 MATERIALS AND METHODS
 RESULTS
DISCUSSION
 REFERENCES
 
Results for feed intake, weight gain, and feed:gain ratios are presented in Table 2Go. There were no significant differences in feed intake among the treatments in the periods from 0 to 3 wk and 0 to 6 wk of age. Feed intake for the 0.075% Hemicell group was greater (P < 0.05) than the other groups from 4 to 6 wk. Significant difference in weight gain among the treatments was not observed from 0 to 3 wk of age. Supplementation with 0.025, 0.05, and 0.075% Hemicell improved (P < 0.05) the weight gain, respectively, by 3.51, 5.06, and 5.39% from 3 to 6 wk and by 2.86, 4.64, and 3.18% from 0 to 6 wk of age. Feed conversion ratio for the 0.025 and 0.05% groups was significantly better (P < 0.05) than in the control group from 4 to 6 wk (3.72 and 4.96%) and from 0 to 6 wk (2.14 and 5.07%). Broilers receiving 0.05% Hemicell yielded better weight gain and feed conversion ratio than the other groups.


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  		Table 2. Effect of Hemicell on growth performance of broilers
 
Effects of Hemicell on relative immune organ weights are presented in Table 3Go. Supplemental Hemicell improved the relative weights of spleen and bursa compared with the control group at wk 3 of age. Relative spleen weights of broilers provided with 0.025% Hemicell were significantly greater than those with higher rates of Hemicell addition and the control group. Regardless of its level, Hemicell supplementation significantly increased relative weight of the bursa compared with the negative control group. In 6-wk-old broilers, supplementation of 0.025, 0.05, and 0.075% Hemicell increased (P < 0.05) the relative weight of the thymus by 22.6, 28.4, and 18.9%, respectively. There were no significant differences in relative immune organ weights among different Hemicell levels. Relative bursa weight of the 0.075% Hemicell group was greater (P < 0.05) than that of other groups.


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  		Table 3. Effect of Hemicell on relative immune organ weights of broilers
 
Effects of Hemicell on serum Ig concentration are presented in Table 4Go. There were no significant differences in IgA and IgG concentrations among the treatments. The best response to the enzyme supplementation occurred in birds receiving 0.05% Hemicell. Compared with the control group, supplementation with 0.025, 0.05, and 0.075% Hemicell significantly increased (P < 0.05) the concentration of IgM produced in wk 3 by 17.2, 6.57, and 9.02%, respectively. Serum IgM concentrations in 6-wk-old broilers provided with 0.05 and 0.075% Hemicell were greater (P < 0.05) than in the other 2 groups.


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  		Table 4. Effect of Hemicell on the concentration of serum immunoglobulin of broilers
 
Effects of Hemicell on T-lymphocyte proliferation are presented in Table 5Go. Proliferation of T lymphocytes in 6-wk-old broilers supplemented with 0.05% Hemicell and the control group were tested and was significantly improved (P < 0.05) in birds provided with 0.05% Hemicell compared with the control group.


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  		Table 5. Effect of Hemicell on T-lymphocyte proliferation
 

   DISCUSSION
TOP
 ABSTRACT
 INTRODUCTION
 MATERIALS AND METHODS
 RESULTS
 DISCUSSION
REFERENCES
 
Soybean meal contains ß-mannan, which can decrease utilization of the feed; ß-mannanase addition could improve utilization of soybean meal. In this experiment, addition of Hemicell improved the average weight gain and feed conversion ratio of broilers. The results were consistent with the findings of McNaughton et al. (1998) and Jackson et al. (2004) who reported that Hemicell improved weight gain and feed conversion ratio of broilers. Addition of 0.05% Hemicell significantly increased the average weight gain of broilers; there was no significant difference among the 0.025, 0.05, and 0.075% Hemicell groups. Jackson et al. (2004) reported inclusion of ß-mannanase at 80 million U/ton improved broiler gains, whereas increasing the inclusion level to 110 million U/ton resulted in no significant additional response. Lee et al. (2005) reported that Hemicell significantly reduced the feed:gain ratio of guar germ diets to a level comparable with the positive and negative control diets, whereas this experiment indicates that different levels of Hemicell addition reduced the feed:gain ratio. Daskiran et al. (2004) added Hemicell at 0.5, 1, and 1.5% in a corn-soy-based starter diet containing 1% guar gum, and found that Hemicell improved feed:gain ratio at all inclusion levels. The current experiment also indicated that Hemicell improved feed:gain ratio at all inclusion levels. Experiments have demonstrated reductions in insulin secretion associated with ß-mannan intake in swine (Sambrook and Rainbird, 1985) and reduced glucose absorption in swine (Rainbird et al., 1984). Addition of an amount of Hemicell may improve insulin secretion and glucose absorption by hydrolyzing ß-mannan. In this experiment, addition of 0.05% Hemicell resulted in the best effect (0.025 to 0.05% is the manufacturer’s recommendation).

In this experiment, Hemicell supplementation increased relative immune organ weights (except for relative thymus weight of 3-wk-old broilers provided with 0.075% Hemicell and relative bursa weight of 6-wk-old broilers provided with 0.025% Hemicell). Supplementation also increased the concentration of serum IgM and T-lymphocyte proliferation of 6-wk-old broilers provided with 0.05% Hemicell. The series of results show that Hemicell may improve the immunity of broilers. The significant effect of Hemicell on immunity of broilers may be explained by the findings of Wu et al. (2005) who reported that substrate of Hemicell entering the intestinal tract resulted in a reduction of the ß-mannan content associated with a reduction of innate immune stimulation. One possible reason why ß-mannanase might improve immunity is that ß-mannan is degraded to mannan oligosaccharide (MOS). Huang et al. (2003) also reported that ß-mannanase hydrolyzed ß-mannan in soybean meal to MOS. Mannan oligosaccharide could influence the immune system, which had been observed by Shashidhara and Devegowda (2003), who reported that MOS significantly increased maternal antibody levels of broilers. Mannan oligosaccharide may also improve the intestinal absorption of some nutrients such as Zn, Cu, and Se (Shao et al., 2000).

In conclusion, the addition of Hemicell improved average weight gain and feed conversion of broilers from 4 to 6 and 0 to 6 wk of age. Hemicell supplementation increased most of the relative immune organ weights and significantly increased the concentration of serum IgM. Hemicell supplementation at 0.05% significantly increased T-lymphocyte proliferation.


   ACKNOWLEDGMENTS
 
The authors gratefully acknowledge C. H. Hu for her skilled technical assistance. The financial provided by Zhejiang Science and Technology Office (Project 2005C22G2010012) is gratefully acknowledged.

Received for publication December 11, 2005. Accepted for publication July 26, 2006.


   REFERENCES
TOP
 ABSTRACT
 INTRODUCTION
 MATERIALS AND METHODS
 RESULTS
 DISCUSSION
 REFERENCES
 
AOAC. 1990. Official Methods of Analysis. 15th ed. Association of Official Analytical Chemists, Arlington, VA.

Bendich, A. P., S. Apolito, E. Gabriel, and L. J. Machlin. 1984. Interaction of dietary vitamin C and vitamin E on Guinea pig immune response to mitogens. J. Nutr. 114:1588–1597.[Abstract/Free Full Text]

Daskiran, M., R. G. Teeter, D. W. Fodge, and H. Y. Hsiao. 2004. An evaluation of endo-ß-D-mannanase (Hemicell) effects on broiler performance and energy use in diets varying in ß-mannan content. Poult. Sci. 83:662–668.[Abstract/Free Full Text]

Huang, X. W., X. S. Liu, and F. Q. Xu. 2003. The effect of ß-mannanase on growth performance of growing pigs. Feed Res. 2003:29–31.

Jackson, M. E., K. Geronian, A. Knox, J. McNab, and E. McCartney. 2004. A dose-response study with the feed enzyme ß-mannanase in broilers provided with corn-soybean meal based diets in the absence of antibiotic. Poult. Sci. 83:1992–1996.[Abstract/Free Full Text]

Lee, J. T., C. A. Bailey, and A. L. Cartwright. 2003. ß-Mannanase ameliorates viscosity-associated depression of growth in broiler chickens fed guar germ and hull fractions. Poult. Sci. 82:1925–1931.[Abstract/Free Full Text]

Lee, J. T., S. Connor-Appleton, C. A. Bailey, and A. L. Cartwright. 2005. Effect of guar meal by-product with and without ß-mannanase Hemicell on broiler performance. Poult. Sci. 84:1261–1267.[Abstract/Free Full Text]

McNaughton, J. L., H. Hsiao, D. A. Maddenn, and D. W. Fodge. 1998. Corn/soy/fat diets for broilers, ß-mannanase and improved feed conversion. Poult. Sci. 77(Suppl. 1):153. (Abstr.)

NRC. 1994. Nutrient Requirements of Poultry. 9th rev. ed. Natl. Acad. Press, Washington, DC.

Patel, M. B., and J. McGinnis. 1985. The effect of autoclaving and enzyme supplementation of guar meal on the performance of chicks and laying hens. Poult. Sci. 64:1148–1156.[ISI][Medline]

Petty, L. A., S. D. Carter, and B. W. Senne. 2002. Effect of beta-mannanase addition to corn-soybean meal diets on growth performance, carcass traits and nutrient digestibility of weanling and growing-finishing pigs. J. Anim. Sci. 80:1012–1019.[Abstract/Free Full Text]

Rainbird, A. L., A. G. Low, and T. Zebrowska. 1984. Effect of guar gum on glucose and water absorption from isolated loops of jejunum in conscious growing pigs. Br. J. Nutr. 52:489–498.[ISI][Medline]

Sambrook, I. E., and A. L. Rainbird. 1985. The effect of guar gum and level and source of dietary fat on glucose tolerance in growing pigs. Br. J. Nutr. 54:27–35.[ISI][Medline]

Shao, L. P., L. J. Zhou, and Z. F. Fei. 2000. Effect of mannan oligosaccharide on immune function and blood antioxidant enzymes in sucking piglets. Acta Nutr. Sinica 22:82–84.

Shashidhara, R. G., and G. Devegowda. 2003. Effect of dietary mannan oligosaccharide on broiler breeder production traits and immunity. Poult. Sci. 82:1319–1323.[Abstract/Free Full Text]

Wu, G., M. M. Bryant, R. A. Voitle, and D. A. Roland, Sr. 2005. Effect of ß-mannanase in corn-soy diets on commercial leghorns in second-cycle hens. Poult. Sci. 84:894–897.[Abstract/Free Full Text]





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