BioPlus® YC produces NSP enzymes in feed assays

A H N H ighlight M 1 007-1 v s1
M ode of a ction
BioPlus® YC produces NSP enzymes in feed assays
Pigs and poultry cannot produce NSP enzymes, and
supplementing microbes that can synthesize NSP
enzymes improve the nutritional value of feed
ingredients. NSP enzyme activity can be analyzed for
each single enzyme (i.e. endo-cellulases) or by the
measurement of the degradation products when fiber is
digested: reducing sugars. A reducing sugar is any
sugar that either has an aldehyde group or is capable of
forming one to allow the sugar to act as a reducing
agent. Reducing sugars include glucose, glyceraldehyde
and galactose as well as disaccharides, such as lactose
and maltose.
This paper presents an in vitro feed assay quantifying
the effect of inoculating feed with BioPlus® Bacillus
strains on reducing sugar content.
Materials and Methods
Compound feed from a US grower finisher farm was
autoclaved at 1210 C for 15 minutes to kill other
microorganisms. Then the feed sample was diluted with
sodium phosphate buffer to ensure a pH of 6-6.5
throughout the experiment. Bacillus products were
added at a dosage rate suggested for finisher pigs (Table
1). A sample was taken for colony forming units (cfu)
determination and analyses for reducing sugar (DNS)
(T=24). The sample was incubated at 370C for 24 hours
and cfu were analyzed. The sample was then centrifuged
and the supernatant was used for DNS estimation.
Reducing sugar was analyzed by 3.5-dinitrosalicylic acid
(DNS) assay described by Gusakov et al. (2012). DNS is
an aromatic compound that reacts with reducing sugars
and other reducing molecules to form 3-amino-5nitrosalicylic acid, which absorbs light strongly at 540
nm.
Acetate buffer was mixed with the Bacillus sample
supernatant and incubated at 40°C for 10 minutes. DNS
reagent was added to the test tube, mixed and
incubated in a boiling water bath for five minutes. After
cooling, absorbance was measured at 540nm in a
spectrophotometer. A standard curve was established
for presenting results in enzyme units (amount of
enzyme needed to release one µmol glucose equivalent
in one ml in one minute). Results in this paper are
presented in optical density (OD) units.
Table 1: Expected dosages of Bacillus products used in the
assay
BioPlusYC
Calsporin
cfu/g feed
6
1.28 x 10
5
3 x 10
g/ton feed
400
30
Results and Conclusion
All Bacillus products supply more nutrients to the animal
by delivering more reducing sugar than the feed sample
without Bacillus (Figure 1, Table 1). BioPlus® YC delivers
about 3 times more reducing sugars than the control.
The competitor, Calsporin, delivers less than Bioplus
YC, less than twice the control.
Figure 1: Effect of Bacillus on reducing sugar in feed (mean of 4
replicates) (T0=time 0; T24=after 24 hr)
Optical Density
Background
Increasing raw material prices are a challenge for animal
production worldwide. Feed prices account for about
70% of pig production cost. However, feed utilization
in the animal is not optimal and a proportion of the feed
ration (15-25%) is not digested by pigs due to a lack of
fiber or non-starch polysaccharide (NSP) degrading
enzymes in the animal’s intestine (Barletta, 2011). An
increase in fiber digestibility increases the availability of
other nutrients that have been locked in by the fiber
matrix. This results in the fiber itself supplying energy,
resulting in reduced nutrient loss to the environment.
4 2 0 BioPlusYC
Calsporin
T0
Control
T24
Table 1: Effect of Bacillus on reducing sugar in feed (OD, mean
± STD)
T0
T24
BioPlus® YC
0.37 ± 0.05
3.53 ± 1.43
Calsporin
0.40 ± 0.11
2.44 ± 0.91
Control
0.41 ± 0.18
1.29 ± 0.77
References
Barletta, 2011. Introduction: Current Market and Expected Developments.
In “Enzymes in Farm Animal Nutrition”, CABI, UK, 2 n d ed. 1-11.
Gusakov, A.V, Kondratyeva, E.G., Sinitsyn, A.P. 2011. Comparison of
two methods for assaying reducing sugars in the determination of
carbohydrase activities. International Journal of Analytical Chemistry.
Article ID 283658. 4 pp.
Project no. 2PROJ1001403. Prepared by AHN Innovation

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