Supplementation of b-mannanase to starter

and grower diets for broilers

C. Kong1, J. H. Lee2, and O. Adeola1,3
1
Department of Animal Sciences, Purdue University, West Lafayette, IN 47907, USA; and 2CTCBIO Inc., Seoul,
Korea. Received 26 July 2010, accepted 8 March 2010.
Kong, C., Lee, J. H. and Adeola, O. 2011. Supplementation of b-mannanase to starter and grower diets for broilers. Can. J.
Anim. Sci. 91: 389397. Two experiments were conducted to investigate the efficacy of b-mannanase on ileal nutrient
digestibility, total tract utilization of dry matter (DM), N, energy, and apparent metabolizable energy (AME, exp. 1), and
growth performance (exp. 2) of birds fed practical corn-soybean meal (SBM)-based diets. In each experiment, 192 male
broilers were assigned to four diets arranged in a 22 factorial of energy level [corn-SBM-based diet that met or exceeded
NRC nutrient requirements (AE) or low energy (LE) diet containing 100 kcal of ME kg1 less than the AE diet] and
Can. J. Anim. Sci. Downloaded from pubs.aic.ca by 202.153.47.194 on 03/05/13

enzyme supplementation (with or without b-mannanase) for 21 d. Supplementing the diet with b-mannanase increased the
birds’ apparent ileal DM digestibility of the experimental diets (PB0.05), whereas there was no effect of energy level.
Neither b-mannanase supplementation nor energy level had any effect on apparent ileal digestibility of any of the amino
acids. The interaction between enzyme and energy levels was not significant for any criteria measured in the excreta except
for DM (P0.05). Addition of b-mannanase improved (PB0.01) the apparent total tract utilization of DM and energy in
the broilers, and there was the same trend (P0.06) for N utilization. Supplementing the diet with b-mannanase also
improved the AME and AMEn of diets by 4.6 and 5.0%, respectively. For all growth performance responses in exp. 2, the
interaction between enzyme and energy levels was not significant. During the starter period (day 2 to 22 post-hatch),
b-mannanase supplementation significantly (PB0.05) increased the body weight (BW) gain of birds. During the grower
period (day 22 to 44 post-hatch) and for the overall period there was no significant difference in BW gain or feed intake of
birds regardless of energy level and b-mannanase supplementation. There were no dietary effects on feed efficiency of birds
during the starter and the grower periods, whereas the birds fed AE diets had higher feed efficiency (PB0.01) than the birds
For personal use only.

fed LE diets. The results indicate that supplementing with b-mannanase may improve gain and energy utilization of
broilers, whereas it may not affect ileal amino acid digestibility.

Key words: Broiler chickens, growth performance, enzyme, b-mannanase, metabolizable energy

Kong, C., Lee, J. H. et Adeola, O. 2011. Enrichissement de la ration de démarrage et de croissance des poulets de chair avec
du b-mannanase. Can. J. Anim. Sci. 91: 389397. Les auteurs ont procédé à deux expériences afin de vérifier l’efficacité de la
b-mannanase sur la digestibilité des éléments nutritifs dans l’iléon, sur l’assimilation de la matière sèche (MS), du N, de
l’énergie et de l’énergie métabolisable apparente (EMA, première expérience) par le tube digestif), ainsi que sur la
croissance (deuxième expérience) des oiseaux nourris avec une ration à base de maı̈s et de tourteau de soja (TS). Dans
chaque expérience, ils ont réparti 192 poulets de chair mâles entre quatre régimes disposés de manière factorielle 22 en
fonction de la concentration d’énergie [ration de maı̈s-TS égale ou supérieure aux besoins énergétiques recommandés par le
NRC (AE) ou ration à faible teneur énergétique (LE) contenant 100 kcal d’énergie métabolisable par kilo de moins que la
ration AE] et de l’enrichissement ou pas avec un enzyme (b-mannanase). Les oiseaux ont été nourris pendant 21 jours.
Enrichir la ration avec du b-mannanase améliore la digestibilité de la MS des rations expérimentales dans l’iléon (PB0,05),
mais la teneur en énergie n’a aucune incidence sur ce plan. Ni le supplément de b-mannanase ni la teneur en énergie n’ont
d’effet sur la digestibilité apparente des acides aminés dans l’iléon. L’interaction de l’enzyme avec la teneur en énergie n’est
pas significative pour les propriétés évaluées au moyen des excréments, hormis la MS (P0,05). L’addition de b-
mannanase rehausse (PB0,01) l’assimilation apparente de la MS et de l’énergie dans le tube digestif des poulets de chair, et
on observe la même tendance (P0,06) pour l’assimilation du N. Enrichir la ration avec du b-mannanase améliore aussi
l’EMA et l’EMAn des rations respectivement de 4,6 et de 5,0%. L’interaction de l’enzyme avec la teneur en énergie n’est
significative pour aucun des facteurs de croissance examinés dans la deuxième expérience. Lors de la période de démarrage
(du 2e au 22e jour suivant l’éclosion), le supplément de b-mannanase a sensiblement (PB0,05a) accru le gain de poids
corporel des volatiles. Pendant la période de croissance (22e au 44e jour suivant l’éclosion) et pour toute la durée de l’étude,
les auteurs n’ont pas observé de variation significative au niveau du gain de poids corporel ni de l’ingestion des aliments,
peu importe la teneur en énergie et la présence ou pas de b-mannanase. La ration n’a eu aucun effet sur la valorisation des
aliments chez les oiseaux durant la période de démarrage et la période de croissance, mais ceux nourris avec la ration AE

Abbreviations: AE, corn-SBM-based diet that met or exceeded

NRC nutrient requirements; AME, apparent metabolizable energy;
AMEn, nitrogen corrected apparent metabolizable energy; BW,
body weight; DDGS, distillers’ dried grains with solubles; DM, dry
matter; LE, diet containing 100 kcal of ME kg1 less than the AE
3
Corresponding author: (e-mail: ladeola@purdue.edu). diet; NSP, nonstarch polysaccharides; SBM, soybean meal

Can. J. Anim. Sci. (2011) 91: 389397 doi:10.4141/CJAS10066 389

 390 CANADIAN JOURNAL OF ANIMAL SCIENCE
valorisaient mieux les aliments (PB0,01) que ceux recevant la ration LE. Ces résultats indiquent que l’addition de
b-mannanase pourrait améliorer le gain de poids et l’assimilation de l’énergie chez les poulets de chair sans toutefois
affecter la digestibilité des acides aminés dans l’iléon.
Mots clés: Poulets de chair, croissance, enzyme, b-mannanase, énergie métabolisable
The identification and reduction of dietary anti-
nutritional factors are necessary for successful poultry
production. It is realized that nonstarch polysaccharides
(NSP) are anti-nutritive through adverse effects on the
digestion and absorption of other nutrients. The term
NSP has been widely adopted in the nutrition field to
cover all polysaccharide molecules excluding starch
(Choct 2002). Among NSP, b-mannans are a group
of related heat-resistant compounds that survive the
Can. J. Anim. Sci. Downloaded from pubs.aic.ca by 202.153.47.194 on 03/05/13
drying-toasting phase of processing soybeans (Dale
1997), and make up to 1.3 or 1.6% in the dehulled or
nondehulled soybean meal (SBM), respectively (Hsiao
et al. 2006). b-Mannans, are mainly found in the hull
and fiber fractions of SBM (Reid 1985), and are highly
viscous and intensely anti-nutritional. The high viscosity
of b-mannans decreases gain:feed ratio and the effi-
ciency for carbohydrate utilization of non-ruminant
animals by partially limiting the access of digestive
enzymes to substrates (Dale 1997).
For personal use only.
One of the common NSP found in corn distillers’
dried grains with soluble (DDGS) and soy-hulls is
b-mannan (Tucker et al. 2004; Hsiao et al. 2006).
Non-dehulled SBM contains 5 to 6% soy-hulls, and
SBM has been widely used in broiler diets as a protein
source. Lumpkins et al. (2004) indicated that corn-
DDGS from modern ethanol plants can be safely used
at 6% in broiler starter diets and 12 to 15% in grower
and finisher diets. Therefore corn-DDGS and soy-hulls
could be potential sources of b-mannan in poultry diets
and b-mannanase supplementation may beneficially
affect performance and nutrient utilization of broilers
fed diets including corn-DDGS and soy-hulls.
It is well documented that enzyme supplementation
in poultry diets can improve efficiency of converting
low-quality protein feedstuffs into high-quality broiler
meat protein for humans (Annison and Choct 1991;
Campbell and Bedford 1992; Bedford and Morgan
1996). Positive effects of bacterial b-mannanase supple-
mentation of poultry diet with copra and guar meals,
feedstuffs rich in b-mannans, have been reported
(Verma and McNab 1982; Patel and McGinnis 1985;
Teves et al. 1988). Daskiran et al. (2004) also reported
that endo-b-mannanase improved feed:gain, water:feed
ratio, and total dry fecal output of broilers by degrading
b-mannans. Jackson et al. (2004) demonstrated that
b-mannanase inclusion at 80 million units per tonne
improved both body weight (BW) gains and feed
conversion of broilers fed corn-SBM based diets.
However, there is a dearth of studies on the effect of
b-mannanase supplementation to corn-SBM diets con-
taining corn-DDGS and soy-hulls on performance,
nutrient digestibility and energy utilization in broilers.
Thus, the objective of this experiment is to determine the
efficacy of a commercial b-mannanase in improving
growth performance, nutrient digestibility and energy
utilization in broilers fed practical corn-SBM diets
containing soy-hulls and corn-DDGS.
MATERIALS AND METHODS
Feeding and collection protocols in the present study
were approved by Purdue University Animal Care and
Use Committee.
General Procedures
Two experiments were conducted to evaluate the efficacy
of a commercial b-mannanase product (CTCZYME† ,
CTCBIO Inc., Seoul, Korea) on growth performance,
apparent ileal digestibility, apparent total tract utiliza-
tion of nutrients, apparent metabolizable energy (AME)
and nitrogen corrected apparent metabolizable energy
(AMEn) of broilers fed corn-SBM based diets with or
without soy-hulls and corn-DDGS. The enzyme product
was produced to have a b-mannanase activity of 800
IU g1. One IU is defined as the quantity of enzyme
required to generate 1 mmol of reducing sugar min 1, at
pH 6.0 and 508C (Miller 1959). The gene source of the
b-mannanase enzyme is a Bacillus subtilis WL-1-derived
Endo1-4 b-mannanase. Three hundred and eighty four 1-
d-old male broilers (Ross 308) were obtained from a local
hatchery. For each experiment, 192 birds were tagged,
individually weighed, allocated into six blocks by BW,
and randomly allotted to four dietary treatments within
a block with eight birds per cage (exp. 1, 0.35 m2) or floor
pen (exp. 2, 3.72 m2) in a randomized complete block
design using the Experimental Animal Allotment Pro-
gram of Kim and Lindemann (2007). Light was provided
for 24 h. Birds were given ad libitum access to water and
diets in both experiments. Experimental treatments
consisting of two factors in a 22 factorial arrangement
of energy and enzyme levels (CTCZYME† ) were as
follows: (1) Adequate energy (AE): A standard corn-
SBM diet that met or exceeded National Research
Council (NRC 1994) nutrient requirements of broilers.
(2) Low energy (LE): As (1) with a decrease of 100 kcal of
ME kg 1 of feed using soyhull and corn-DDGS at the
expense of corn and SBM. (3) AECTCZYME† : As
(1) with 400 IU of b-mannanase kg 1 of feed. (4) LE
CTCZYME† : As (2) with 400 IU of b-mannanase kg1
of feed.
 KONG ET AL. * b-MANNANASE SUPPLEMENTATION TO BROILER DIET
Experiment 1
In this experiment, energy and nutrient utilization
responses to b-mannanase over a 3-wk period from
days 1 to 22 post-hatch were evaluated. The birds were
housed in electrically heated battery cages (Alternative
Design Manufacturing, Siloam Spring, AR 72761,
model # SB 4 T) in an environmentally controlled
room. Battery temperature on days 18, 815, and 1522
post-hatch were kept at 35, 32 and 278C, respectively.
Chromic oxide was added as an indigestible marker
to enable the determination of nutrient digestibility of
diets using the index method (Kong and Adeola 2010).
Sample Collection and Analyses
Can. J. Anim. Sci. Downloaded from pubs.aic.ca by 202.153.47.194 on 03/05/13
Excreta samples were collected twice daily on days 19,
20, and 21 post-hatch. During collection, waxed paper
was placed in trays under the cages and excreta on the
paper were collected. The excreta samples were pooled
per cage over the 3 d, and stored in a freezer. On day
22 post-hatch, birds were asphyxiated with CO2, the
distal part of ileum (portion of the small intestine from
Meckel’s diverticulum to approximately 1 cm anterior to
the ileo-cecal junction) was removed, and the contents
were gently rinsed with distilled water into plastic
containers and stored frozen at 208C. Excreta were
For personal use only.
dried in an oven at 558C for 5 d (Adeola et al. 2008) and
ground to pass through a 0.5-mm screen using a mill
grinder (Retsch ZM 100, GmbH & Co. K.C., Germany).
All ileal digesta samples were subsequently freeze-dried
and ground using a coffee grinder.
Duplicate proximate analyses were performed on
diets, excreta, and ileal digesta samples. Dry matter
analysis of samples was conducted by drying the
samples in a drying oven at 1058C for 24 h [method
934.01, Association of Official Analytical Chemists
(AOAC) 2006]. Subsamples of the diets, excreta, and
ileal digesta were analyzed for nitrogen using the
combustion method (Model FP2000, LECO Corp, St.
Joseph, MI). The gross energy of the samples was deter-
mined by adiabatic bomb calorimeter (Parr 1261 bomb
calorimeter, Parr Instruments Co., Moline, IL) using
benzoic acid as a calibration standard. The diets and
ileal digesta samples were analyzed for amino acids.
Samples for amino acid analysis were prepared using a
24-h hydrolysis in 6 N hydrochloric acid at 1108C under
an atmosphere of nitrogen. For methionine and cy-
steine, performic acid oxidation was done prior to acid
hydrolysis. Samples for tryptophan analysis were hydro-
lyzed using barium hydroxide. Amino acids in hydro-
lysates were determined by HPLC after post-column
derivatization. All amino acid analyses were done at the
University of Missouri Experiment Station Chemical
Laboratory [method 982.30 E (a, b, c), AOAC (2006)].
Chromium concentration in the diets, ileal, and excreta
samples was determined using the method of Fenton
and Fenton (1979).
391
Calculations
Apparent ileal nutrient digestibility and total tract
utilization (C, %), and AME were calculated using the
following equations (Kong and Adeola 2010):
C [1(Cri =Cro )(No =Ni )]100
AME  GE C=100
Where Cri is the concentration of chromium in the diet,
Cro is the concentration of chromium in the ileal digesta
or excreta, No is the concentration of the nutrient or
energy in the ileal digesta or excreta, and Ni is the
concentration of the nutrient or energy in the diet. All
values for Cri, Cro, No and Ni are expressed as a
percentage of dry matter. The product of C (total tract
utilization) and the gross energy (GE, kcal kg1) of the
diet is AME (kcal kg1) of the diet. Because catabolic
compounds in excreted nitrogen can contribute to
energy loss, metabolizable energy was corrected to
zero nitrogen retention using a factor of 8.22 kcal g1
(Hill and Anderson 1958).
Experiment 2
To further evaluate the efficacy of the b-mannanase in
improving growth performance, exp. 2 was conducted
over a 42-d period from day 2 to day 44 post-hatch. Bird
weights and feed consumption were measured on a pen
basis at days 22 and 44 post-hatch. Mortality was
recorded on a daily basis.
The birds were placed in floor pens (0.465 m2 bird 1)
with fresh wood shavings in an environmentally con-
trolled broiler house. Each pen had a floor feeder and a
bell waterer. The feeding program consisted of a starter
(day 2 to day 22 post-hatch) and a grower (day 22 to day
44 post-hatch) phase. Starter diets were same as exp. 1
diets except for corn, indigestible marker and limestone
(Table 1).
Statistical Analysis
Data were analyzed as a randomized complete block
design using the MIXED procedure of SAS software
(SAS Institute, Inc. 2006). The model for data from exp.
1 and exp. 2 included the two main effects (energy and
enzyme levels) and the interaction as fixed effects, and
block as a random effect. In all analyses, cage (exp. 1) or
pen (exp. 2) was considered the experimental unit and
least squares means were calculated for each variable. If
an interaction was significant (PB0.05), the SLICE
option for the LSMEANS statement was used to
analyze simple effects. A level of PB0.05 was considered
statistically significant.
RESULTS
The results of b-mannanase analysis of diets showed
that b-mannanase activity in diets without added b-
mannanase were less than 100 IU kg1. In exp. 1, the
analyzed b-mannanase activity was 760 and 530 IU kg1
for AE and LE diets with b-mannanase supplementation,
 392 CANADIAN JOURNAL OF ANIMAL SCIENCE

Table 1. Ingredients and chemical composition of the experimental diets

Exp. 1 Exp. 2

Starter Starter Grower

Ingredient (g kg 1) AEz LEz AE LE AE LE

Corn 543.0 499.5 546.0 502.5 606.5 563.0

Soybean meal 352.5 346.0 352.5 346.0 307.5 301.0
Soy hull 0.0 25.0 0.0 25.0 0.0 25.0
DDGS 0.0 25.0 0.0 25.0 0.0 25.0
Soy oil 60.0 60.0 60.0 60.0 50.0 50.0
Dicalcium phosphatey 18.0 18.0 18.0 18.0 14.5 14.5
Limestone 12.0 12.0 14.0 14.0 13.5 13.5
Salt 4.0 4.0 4.0 4.0 4.0 4.0
Chromic oxide 5.0 5.0    
Vitamin-mineral premixx 3.0 3.0 3.0 3.0 3.0 3.0
DL-Methionine 2.5 2.5 2.5 2.5 1.0 1.0
Can. J. Anim. Sci. Downloaded from pubs.aic.ca by 202.153.47.194 on 03/05/13

Total 1000.0 1000.0 1000.0 1000.0 1000.0 1000.0

Calculated composition
Crude protein (g kg1) 220 220 220 220 202 202
ME (kcal kg 1) 3183 3084 3193 3094 3202 3102
Ca (g kg 1) 9.2 9.2 10.0 10.0 9.0 9.0
P (g kg 1) 7.0 7.1 7.0 7.1 6.3 6.3
Non-phytate P (g kg1) 4.5 4.6 4.5 4.6 3.8 3.9
Total amino acids (g kg1)
Arg 14.3 14.2 14.3 14.2 13.0 12.9
His 5.8 5.7 5.8 5.7 5.3 5.3
Ile 9.0 9.0 9.1 9.0 8.3 8.3
Leu 18.6 18.5 18.6 18.5 17.6 17.4
For personal use only.

Lys 11.8 11.7 11.9 11.7 10.7 10.6

Met 5.8 5.9 5.8 5.9 4.2 4.2
Cys 3.5 3.5 3.5 3.5 3.3 3.3
Phe 10.3 10.3 10.3 10.3 9.5 9.5
Tyr 8.5 8.4 8.5 8.4 7.8 7.7
Thr 8.2 8.1 8.2 8.2 7.5 7.5
Trp 2.9 2.9 2.9 2.9 2.6 2.6
Val 10.0 10.0 10.0 10.0 9.3 9.3
Analyzed composition
CP (N6.25) (g kg1) 221 208 218 211 195 198
GE (kcal kg 1) 4562 4607 4686 4740 4604 4629

z
AE, a standard corn-SBM diet that met NRC (1994) nutrients requirements; LE, a diet containing 100 kcal of ME kg1 less than the AE diet.
y
20% Ca, 18.5% P.
x
Supplied the following per kilogram of diet: vitamin A, 5484 IU; vitamin D3, 2643 ICU; vitamin E,11 IU; menadione sodium bisulfite,4.38 mg;
riboflavin, 5.49 mg; D-pantothenic acid, 11 mg; niacin, 44.1 mg; choline chloride, 771 mg; vitamin B12, 13.2 mg; biotin, 55.2 mg; thiamine mononitrate,
2.2 mg; folic acid, 990 mg; pyridoxine hydrochloride, 3.3 mg; I, 1.11 mg; Mn, 66.06 mg; Cu, 4.44 mg; Fe, 44.1 mg; Zn, 44.1 mg; Se, 300 mg.

respectively. In exp. 2, starter diets, 770 and 580 IU kg 1
of b-mannanase activity was analyzed in the AE and LE
diets, respectively. For the b-mannanase-supplemented
grower diets in exp. 2, analyzed activity was 780 and
800 IU kg1 for the AE and LE diets, respectively.
Experiment 1
The main effects of b-mannanase and energy on appar-
ent ileal digestibility of dry matter (DM), nitrogen, and
amino acids for broilers fed the experimental diets with
two levels of energy and b-mannanase in exp. 1 are
presented in Table 2. For all criteria measured at the
ileum, the interaction between enzyme and energy levels
was not significant and, thus, only the main effects means
are presented. Apparent ileal DM digestibility was
increased by b-mannanase supplementation (P 0.02),
but was not affected by energy level. Apparent ileal
digestibility of nitrogen was not influenced by the
enzyme or energy level. Also, apparent ileal digestibility
of all amino acids was not affected by supplementation
of b-mannanase or energy level.
Apparent total tract DM, energy, and N utilization of
broilers fed the experimental diets at different energy
level (AE or LE) without or with added b-mannanase are
presented in Table 3. The interaction between enzyme
and energy levels was significant for DM (P0.05) and
thus simple effects of enzyme are presented in order to
determine the differences between predicted values
across enzyme levels for different levels of energy.
Addition of b-mannanase improved (PB0.01) the ap-
parent total tract DM utilization of broilers fed AE diets
and there was a trend (P0.08) for improvement of
 KONG ET AL. * b-MANNANASE SUPPLEMENTATION TO BROILER DIET 393

Table 2. Main effects of b-mannanase and energy on apparent ileal digestibility of DM, nitrogen, and amino acids for broilers fed the experimental diets
with two levels of energy and b-mannanase in exp. 1

b-mannanase Energy P value

Item Not added added AEz LEz SEy b-mannanase Energy Interaction

Digestibility (%)
DM 74.1 76.5 75.2 75.4 1.2 0.02 0.82 0.49
N 85.9 86.3 86.5 85.8 0.7 0.57 0.34 0.87
Essential amino acids
Arg 92.3 92.3 92.4 92.3 0.3 0.97 0.84 0.53
His 88.7 89.0 88.7 89.0 0.6 0.66 0.65 0.96
Ile 87.6 87.4 87.5 87.5 0.5 0.77 0.98 0.68
Leu 88.9 88.8 88.8 88.9 0.5 0.79 0.75 0.48
Lys 89.7 89.8 89.7 89.8 0.5 0.80 0.91 0.91
Met 94.7 95.1 94.8 94.9 0.3 0.28 0.83 0.13
Phe 89.3 89.0 89.1 89.1 0.5 0.63 0.98 0.52
Can. J. Anim. Sci. Downloaded from pubs.aic.ca by 202.153.47.194 on 03/05/13

Thr 83.5 83.2 83.4 83.3 0.8 0.74 0.85 0.44

Trp 89.6 89.0 89.9 88.8 0.7 0.44 0.14 0.54
Val 85.7 85.7 85.6 85.7 0.7 0.99 0.94 0.53
Nonessential amino acids
Ala 88.2 88.1 88.0 88.2 0.6 0.91 0.74 0.61
Asp 86.1 86.0 86.2 86.0 0.6 0.87 0.71 0.51
Cys 79.8 79.9 79.4 80.3 1.0 0.96 0.46 0.53
Glu 91.5 91.5 91.6 91.5 0.3 0.93 0.85 0.60
Gly 83.3 83.7 83.4 83.6 0.9 0.67 0.81 0.72
Pro 88.2 88.2 88.2 88.3 0.6 0.97 0.88 0.44
Ser 87.3 87.3 87.4 87.3 0.7 0.94 0.85 0.27
Tyr 87.9 88.0 87.7 88.1 0.6 0.81 0.49 0.45
Total AA 88.1 88.1 88.1 88.1 0.6 0.96 0.96 0.55
For personal use only.

nx 12 10 11 11

z
AE, a standard corn-SBM diet that met NRC (1994) nutrients requirements; LE, a diet containing 100 kcal of ME kg1 less than the AE diet.
y
Maximum value of standard error for each main effect.
x
Number of replicate cages of eight birds per cage. Data from two cages were removed due to insufficient sample quantity.

DM utilization in birds fed the LE diet. There was a
significant increase (PB0.01) in energy utilization of
the broilers fed b-mannanase supplemented diets and
numerically increased (P0.06) N retention was found
in birds fed diets with b-mannanase.
Table 4 illustrates the main effects of b-mannanase
and energy on AME and AMEn of broilers fed the ex-
perimental diets with different energy and b-mannanase
levels in exp. 1. Because there was no significant
interaction between enzyme and energy levels, main
effects means are presented. There was a significant
increase in dietary AME from 3588 to 3752 kcal kg1
with b-mannanase supplementation (PB0.01). Similarly,
AMEn of diets increased from 3364 to 3531 kcal kg1
with b-mannanase addition (PB0.01). There was no
significant difference in AME between AE and LE diets,

Table 3. Effects of b-mannanase and energy on apparent total tract DM, N and energy utilization of broilers fed the experimental diets with two levels of
energy and b-mannanase in exp. 1

Diet P value

Not added Added Main effects Simple effects of enzyme

b-mannanase Item AEz LEz AE LE SEy Enzyme Energy Interaction AE LE

Utilization (%)
DM 74.7 76.0 77.9 77.2 0.5 B0.01 0.52 0.05 B0.01 0.08
N 71.0 68.9 72.6 71.1 1.0 0.06 0.07 0.77 0.23 0.11
Energy 78.4 79.4 81.3 80.8 0.4 B0.01 0.55 0.08 B0.01 0.04
nx 6 6 6 6

z
AE, a standard corn-SBM diet that met NRC (1994) nutrients requirements; LE, a diet containing 100 kcal of ME kg1 less than the AE diet.
y
Standard error.
x
Number of replicate cages of eight birds per cage.
 394 CANADIAN JOURNAL OF ANIMAL SCIENCE

Table 4. Main effects of b-mannanase and energy on AME and AMEn on a dry matter basis of broilers fed the experimental diets with different energy
and b-mannanase levels in exp. 1

b-mannanase Energy P value

Item Not added Added AEz LEz SEy Enzyme Energy Interaction
1
Metabolizable energy (kcal kg )
AME 3588 3752 3654 3686 14 B0.01 0.12 0.89
AMEn 3364 3531 3426 3469 13 B0.01 0.03 0.53
nx 12 12 12 12

z
AE, a standard corn-SBM diet that met NRC (1994) nutrients requirements; LE, a diet containing 100 kcal of ME kg1 less than the AE diet.
y
Standard error.
x
Number of replicate pen of eight birds per cage.

whereas difference of AMEn (3426 vs. 3469 kcal kg1) LE diets was 6.7% higher (PB0.05) than that of birds
Can. J. Anim. Sci. Downloaded from pubs.aic.ca by 202.153.47.194 on 03/05/13

between AE and LE diets was found to be significant
(PB0.05).
Experiment 2
Growth performance responses of birds fed experimen-
tal AE or LE diets without or with b-mannanase
supplementation from day 2 to day 44 post-hatch are
presented in Table 5. The average BW of birds at day 2
post-hatch (46 g per bird) was identical for all diets. For
all growth performance responses, the interaction be-
For personal use only.
fed AE diets. During the grower period (day 22 to day
44 post-hatch) and for the overall period (day 2 to
day 44 post-hatch), neither b-mannanase addition nor
energy level had any effect on BW gain and feed intake
of birds. There were no dietary effects on feed efficiency
of birds during the starter or the grower periods,
whereas for the overall period the birds fed AE diets
had a higher feed efficiency (P B0.01) than the birds fed
LE diets.

tween enzyme and energy levels was not significant and
thus only the main effects means are presented. During
the starter period (day 2 to day 22 post-hatch), the BW
gain of birds fed diets containing b-mannanase was
3.5% higher (PB0.05) than that of birds fed diets
without b-mannanase, whereas feed intake of birds fed
DISCUSSION
The present experiment showed positive effect of
b-mannanase on BW gain of birds, apparent total tract
DM and energy utilization, and AME and AMEn of
diets in the starter period. The results were consistent
with the findings of Jackson et al. (2004) and Zou et al.

Table 5. Main effects of b-mannanase and energy on BW, BW gain, feed intake and feed efficiency of broilers fed the experimental diets with two levels of
energy and b-mannanase in exp. 2

b-mannanase Energy P value

z z y
Item Not added Added AE LE SE Enzyme Energy Interaction

BW (g bird 1)
Day 2 46 46 46 46
Day 22 770 795 776 788 10 0.01 0.22 0.11
Day 44 3112 3144 3147 3108 54 0.46 0.38 0.56
BW gain (g bird 1)
Starter 724 749 731 742 9 0.01 0.22 0.11
Grower 2342 2350 2371 2321 50 0.84 0.21 0.80
Overall 3066 3098 3101 3062 55 0.46 0.38 0.55
Feed intake (g bird 1)
Starter 1011 1021 983 1049 23 0.76 0.04 0.92
Grower 3970 3999 3992 3976 74 0.65 0.81 0.69
Overall 4981 5019 4975 5026 81 0.54 0.41 0.65
Feed efficiency (g gain kg 1 feed)
Starter 719 738 747 710 18 0.43 0.14 0.65
Grower 590 587 594 584 4 0.66 0.09 0.84
Overall 616 617 623 610 4 0.73 0.01 0.70
nx 12 12 12 12

z
AE, a standard corn-SBM diet that met NRC (1994) nutrients requirements; LE, a diet containing 100 kcal of ME kg1 less than the AE diet.
y
Standard error.
x
Number of replicate cages of eight birds per pen.
 KONG ET AL. * b-MANNANASE SUPPLEMENTATION TO BROILER DIET
(2006), who reported that supplemented b-mannanase
improved weight gain of broilers fed different level of
b-mannannase with corn-soybean meal based diet.
However, the improved BW gain of birds in the
present study is not supported by the simple hypothesis
that AME content of the diet was increased by the
release of simple sugars. b-Mannans are mainly found in
the hull and fiber fraction of SBM (Reid 1985) as well as
in corn-DDGS (Tucker et al. 2004). Hsiao et al. (2006)
indicated that a broiler starter diet containing 35%
dehulled SBM might contain 0.44% b-mannan. Based
on these data, calculated value of b-mannan content
(approximately 0.64%) of LE diets in the current study
might be slightly higher than that of AE diets (approxi-
mately 0.44%). Simple sugars released from this portion
Can. J. Anim. Sci. Downloaded from pubs.aic.ca by 202.153.47.194 on 03/05/13
would not be enough to account for the improved
performance in the present study. b-Mannanase supple-
mentation indeed improved the AME and AMEn of
diets (PB0.01) used in the current study by 4.6 and
5.0%, respectively, which were higher than expected
values that resulted from released carbohydrates by
b-mannanase supplementation. Moreover, there was no
significant change in feed intake or feed efficiency with
the enzyme, which would have been observed if the
hypothesis was true.
In the current study, although AE diets was formu-
For personal use only.
lated to have 100 kcal more ME than that of LE diets,
this did not result in any detectable effects on AME,
whereas difference of AMEn (3426 vs. 3469 kcal kg1)
between AE and LE diets was found to be significant.
These results are difficult to explain, but we assume that
these could, in part be responsible for the lack of
performance difference in starter phase in exp. 2.
The mode of action of b-mannanase includes im-
provement in the reduction of digesta viscosity, the
production of mannanoligosaccharides, the reduction of
innate immune stimulation (Johnson and Gee 1986;
Ross et al. 2002), and improved energy metabolism
(Radcliffe et al. 1999). It is well known that b-mannans
have adverse effects on the digestive system, mainly due
to their highly viscous nature. This highly viscous nature
results in slowing gastric emptying, impairing the mixing
of substrate with digestive enzymes, and reducing the
rate of contact of nutrients with absorptive epithelium
(Read 1986). Maisonnier et al. (2003) reported that
0.5% of guar gum [0.67% of b-mannan in the diet;
calculation based on Hsiao et al. (2006)] supplementa-
tion significantly increased intestinal supernatant visc-
osity and decreased BW gain of broilers from day 7 to
day 21. These adverse effects may be overcome with
b-mannanase supplementation. Daskiran et al. (2004)
reported that water consumption per unit of feed
consumption increased with increasing dietary guar
gum, and this was caused by the need to maintain
proper mixing of digestive enzymes with substrates when
guar gum was added to the diet; however, enzyme
supplementation tended to reduce water consumption,
thus counteracting the effect of guar gum. Lee et al.
395
(2003) showed that supplementation of feeds contain-
ing guar meal with b-mannanase reduced intestinal
viscosity and increased weight gain of broiler chicks.
In the current study, increased BW gain (PB0.05),
apparent ileal DM digestibility (PB0.05), and apparent
total tract DM and energy retentions (PB0.01) in the
diets with b-mannanase supplementation for starter
period were observed, which in part, likely resulted
from a b-mannanase-induced reduction in intestinal
viscosity.
In the current study, endo b-1, 4-mannanase improved
broiler BW gain during the starter phase. Because endo
b-1,4-mannanase cleaves randomly within the 1,4-b-D
mannan main chain of galactomannan, glucomannan,
galactoglucomannan, and mannan (McCleary and
Matheson 1986), it can be speculated that mannanoligo-
saccharides, mannotriose and mannobiose, as well as a
small amount of mannose, were generated when endo
b-1,4-mannanase was added to the diet. Because man-
nose can only be absorbed in the small intestine, the
other metabolites cannot supply energy to the chicks.
This supports the concept that the amount of absorbed
simple sugars in the small intestine was not enough to
account for improvement of performance in the present
study. Production of mannanoligosaccharides could
improve chicken health, either by interfering with
bacterial attachment to the epithelial cell (Spring et al.
2000) or by enhancing the immune system (Newman and
Newman 2001; O’Quinn et al. 2001), which may partially
contribute to the improvement in BW gain in diets with
b-mannanase for starter period (Sundu et al. 2006).
Beta-galactomannan has been shown to inhibit secre-
tion of insulin in humans (Morgan et al. 1985) and swine
(Leeds et al. 1980; Sambrook and Rainbird 1985). It has
also been observed that b-mannans reduce glucose and
water absorption in swine (Rainbird et al. 1984). In the
current study, b-mannanase may have elicited improve-
ments in apparent ileal DM digestibility, apparent total
tract DM and energy utilization, and AME through
degradation of b-mannans in the diets. In addition to
this, it is also speculated that there may be an indirect
effect on regulatory hormones but further research is
needed to elucidate this indirect effect.
This study shows that the addition of b-mannanase
improved BW gain, apparent ileal DM digestibility, and
apparent total tract DM and energy utilization of
broilers in the starter period. The addition of b-
mannanase also improved the AME and AMEn of
corn-SBM-based diets used in the current study. These
positive effects of b-mannanase supplementation may
partly result from increased energy contents by degrad-
ing b-mannans in the diets. However, further research is
needed to explain these positive effects by examining
potential mechanisms of b-mannanase, which include
reducing digesta viscosity and innate immune stimula-
tion, and effecting regulatory hormones.
 396 CANADIAN JOURNAL OF ANIMAL SCIENCE
Adeola, O., Shafer, D. J. and Nyachoti, C. M. 2008. Nutrient
and energy utilization in enzyme-supplemented starter
and grower diets for White Pekin Ducks. Poult. Sci. 87:
255263.
Annison, G. and Choct, M. 1991. Antinutritional activities
of cereal non-starch polysaccharides in broiler diets and
strategies minimizing their effects, World. s Poult. Sci. J. 47:
232241.
Association of Official Analytical Chemists. 2006. Official
methods of analysis. 18th ed. AOAC, Washington, DC.
Bedford, M. R. and Morgan, A. J. 1996. The use of enzymes in
poultry diets. World’s Poult. Sci. J. 52: 6168.
Campbell, G. L. and Bedford, M. R. 1992. Enzyme applications
for monogastric feeds: A review. Can. J. Anim. Sci. 72:
449466.
Choct, M. 2002. Non-starch polysaccharides: Effect on nu-
Can. J. Anim. Sci. Downloaded from pubs.aic.ca by 202.153.47.194 on 03/05/13
tritive value. Pages 221235 in J. McNab and N. Boorman,
eds. Poultry feedstuffs: Supply, composition and nutritive
value. CABI Publishing, New York, NY.
Dale, N. 1997. Current status of feed enzymes for swine.
Hemicell, poultry and swine feed enzyme. ChemGen Crop,
Gaithersburg, MD.
Daskiran, M., Teeter, R. G., Fodge, D. W. and Hsiao, H. Y.
2004. An evaluation of endo-b-D-mannanase (Hemicell)
effects on broiler performance and energy use in diets varying
in b-mannan content. Poult. Sci. 83: 662668.
Fenton, T. W. and Fenton, M. 1979. An improved procedure
for the determination of dietary chromic oxide in feed and
For personal use only.
feces. Can. J. Anim. Sci. 59: 631634.
Hill, F. W. and Anderson, D. L. 1958. Comparison of
metabolizable energy and productive determinations with
growing chicks. J. Nutr. 64: 587603.
Hsiao, H. Y., Anderson, D. M. and Dale, N. M. 2006. Levels of
b-mannan in soybean meal. Poult. Sci. 85: 14301432.
Jackson, M. E., Geronian, K., Knox, A., McNab, J. and
McCartney, E. 2004. A dose-response study with the feed
enzyme b-mannanase in broilers provided with corn-soybean
meal based diets in the absence of antibiotic. Poult. Sci. 83:
19921996.
Johnson, I. T. and Gee, J. M. 1986. Gastrointestinal adapta-
tion in response to soluble non-available polysaccharides in the
rat. Br. J. Nutr. 55: 497505.
Kim, B. G. and Lindemann, M. D. 2007. A new spreadsheet
method for the experimental animal allotment. J. Anim. Sci. 85
(Suppl. 2): 112.
Kong, C. and Adeola, O. 2010. Apparent ileal digestibility of
amino acids in feedstuffs for White Pekin ducks. Poult. Sci. 89:
545550.
Lee, J. T., Bailey, C. A. and Cartwright, A. L. 2003.
b-mannanase ameliorates viscosity-associated depression of
growth in broiler chickens fed guar germ and hull fractions.
Poult. Sci. 82: 19251931.
Leeds, A. R., Kang, S. S., Low, A. G. and Sambrook, I. E. 1980.
The pig as a model for studies on the mode of action of guar
gum in normal and diabetic man. Proc. Nutr. Soc. 39: 44.
Lumpkins, B. S., Batal, A. B. and Dale, N. M. 2004. Evaluation
of distillers dried grains with solubles as a feed ingredient for
broilers. Poult. Sci. 83: 18911896.
Maisonnier, S., Gomez, J., Brée, A., Berri, C., Baéza, E. and
Carré, B 2003. Effects of microflora status, dietary bile salts
and guar gum on lipid digestibility, intestinal bile salts, and
histomorphology in broiler chickens. Poult. Sci. 82: 805814.
McCleary, B. V. and Matheson, N. K. 1986. Enzymic analysis
of polysaccharide structure. Adv. Carbohydr. Chem. Biochem.
44: 147176.
Miller, G. L. 1959. Use of dinitrosalicylic acid reagent for
determination of reducing sugar. Anal. Chem. 31: 426428.
Morgan, L. M., Tredger, J. A., Madden, A., Kwasowski, P. and
Marks, V. 1985. The effect of guar gum on carbohydrate, fat
and protein stimulated gut hormone secretion: Modification
of postprandial gastric inhibitory polypeptide and gastrin
responses. Br. J. Nutr. 53: 467475.
Newman, K. E. and Newman, M. C. 2001. Evaluation of
mannan oligosaccharide on the microflora and immunoglobu-
lin status of sows and piglet performance. J. Anim. Sci. 79
(Suppl. 1): 189.
National Research Council. 1994. Nutrient requirements of
poultry. 9th rev. ed. National Academy Press, Washington,
DC.
O’Quinn, P. R., Funderburke, D. W. and Tibetts, G. W. 2001.
Effects of dietary supplementation with mannan oligosacchar-
ides on sow and litter performance in a commercial production
system. J. Anim. Sci. 79 (Suppl. 1): 212.
Patel, M. B. and McGinnis, J. 1985. The effect of autoclaving
and enzyme supplementation of guar meal on the performance
of chicks and laying hens. Poult. Sci. 64: 11481156.
Radcliffe, J. S., Robbins, B. C., Rice, J. P., Pleasant, R. S. and
Kornegay, E. T. 1999. The effects of hemicell on digestibilities
of minerals, energy, and amino acids in pigs fitted with steered
ileo-cecal cannulas and fed a low and high protein corn-
soybean meal diet. J. Anim. Sci. 77 (Suppl. 1): 197. (Abstr.)
Rainbird, A. L., Low, A. G. and Zebrowska, T. 1984. Effect of
guar gum on glucose and water absorption from isolated loops
of jejunum in conscious growing pigs. Br. J. Nutr. 52: 489498.
Read, N. W. 1986. Dietary fiber and bowel transit. Pages 91
100 in G. V. Vahouny and D. Kritchevsky, eds. Dietary fiber
basic and clinical aspects. Plenum Press, New York, NY.
Reid, J. S. G. 1985. Cell wall storage carbohydrate in seeds-
Biochemistry of the seed ‘‘gums’’ and ‘‘hemicelluloses.’’. Adv.
Bot. Res. 2: 125155.
Ross, S. A., Duncan, C. J. G., Pasco, D. S. and Pugh, N. 2002.
Isolation of a galactomannan that enhances macrophage
activation from the edible fungus Morchella esculenta. J.
Agric. Food Chem. 50: 56835685.
Sambrook, I. E. and Rainbird, A. L. 1985. The effects of guar
gum and level and source of dietary fat on glucose tolerance in
growing pigs. Br. J. Nutr. 54: 2735.
SAS Institute, Inc. 2006. SAS/STAT user’s guide. Release 9.1.
SAS Institute Inc., Cary, NC.
Spring, P., Wenk, C., Dawson, K. A. and Newman, K. E. 2000.
The effects of dietary mannanoligosaccharides on cecal para-
meters and the concentrations of enteric bacteria in the ceca of
salmonella-challenged broiler chicks. Poult. Sci. 79: 205211.
Sundu, B., Kumar, A. and Dingle, J. 2006. Response of broiler
chicks fed increasing levels of copra meal and enzymes. Int. J.
Poul. Sci. 5: 1318.
Teves, F. G., Zamora, A. F., Calapardo, M. R. andLuis,
E. S.1988. Nutritional value of copra meal treated with
bacterial mannanase in broiler diets. Pages 497507 in Recent
advances in biotechnology and applied biology. Proceedings of
the Eighth International Conference on Global Impacts of
Applied Microbiology and International Conference on Ap-
plied Biology and Biotechnology. Chinese University Press,
Hong Kong.
 KONG ET AL. * b-MANNANASE SUPPLEMENTATION TO BROILER DIET 397

Tucker, M. P., Nagle, N. J., Jennings, E. W., Ibsen, K. N.,
Aden, A., Nguyen, Q. A., Kim, K. H. and Noll, S. L. 2004.
Conversion of distiller’s grain into fuel alcohol and a higher-
value animal feed by dilute-acid pretreatment. Appl. Biochem.
Biotechnol. 115: 11391159.
Can. J. Anim. Sci. Downloaded from pubs.aic.ca by 202.153.47.194 on 03/05/13
For personal use only.
Verma, S. V. S. and McNab, J. M. 1982. Guar meal in diets for
broiler chickens. Br. Poult. Sci. 23: 95105.
Zou, X. T., Qiao, X. J. and Xu, Z. R. 2006. Effect of b-
mannanase (Hemicell) on growth performance and immunity
of broilers. Poult. Sci. 85: 21762179.