While the inclusion of phytases has become almost ubiquitous in monogastric feeding, both to release phosphorus and to reduce the anti-nutritive effect of phytate itself (superdosing), the use of enzymes to tackle issues associated with non-starch polysaccharides (NSP, i.e. fibre) remains the subject of much discussion. There is still no general consensus on how to achieve the greatest benefits with these enzymes, and the potential to incorporate them into a multi-enzyme dosing strategy has only added to the debate.

1. Digital Re-print -
July | August 2013
Exploring the challenge of single versus multi-
enzyme dosing comparisons
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3. W
hile the inclusion of phytases
has become almost ubiquitous
in monogastric feeding, both
to release phosphorus and to reduce
the anti-nutritive effect of phytate itself
(superdosing), the use of enzymes to
tackle issues associated with non-starch
polysaccharides (NSP, i.e. fibre) remains
the subject of much discussion. There
is still no general consensus on how to
achieve the greatest benefits with these
enzymes, and the potential to incorporate
them into a multi-enzyme dosing strategy
has only added to the debate.
Single vs multiple enzymes
The use of either single commercial
products incorporating multiple enzymes, or
the combination of separate products with
different modes of action, would appear in
theory to have clear advantages, but the
reality is much more complex. As such,
when looking to make commercial decisions
between single versus multi-enzyme dosing,
it is important to be aware of the factors that
influence target animal performance.
The most commonly used commercial
feed enzymes typically fall into one of two
broad categories, namely phytases and NSP
enzymes, with the latter containing a range
of enzymes developed with the aim of
breaking down the various fibre compo-
nents in the diet. This fibrous content will
differ depending on the feed ingredients
used, however, with the main constituents
being cellulose, arabinoxylans, mixed-linked
ß-glucans, glucomannans, galactomannans
and arabinans.
The values in Table 1 illustrate the
Exploring the challenge of single
versus multi-enzyme dosing
comparisons
by Dr Helen Masey O’Neill, research manager, and Tiago dos Santos, global technical
manager, AB Vista, United Kingdom
Table 1: Types and estimated levels of the main fibre polysaccharide components present in key
cereal grains
Cereal
% of dry matter
Xylan1 ß-Glucan Cellulose Mannan2
Galactan3
Uronic
acids4
Total
Wheat Soluble 1.8 0.4 - - 0.2 - 2.4
Insoluble 6.3 0.4 2.0 - 0.1 0.2 9.0
Barley Soluble 0.8 3.6 - - 0.1 - 4.5
Insoluble 7.1 0.7 3.9 0.2 0.1 0.2 12.2
Oats Soluble 1.0 2.8 - 0.2 0.2 0.1 4.3
Insoluble 9.2 0.5 8.2 0.1 0.4 0.7 19.1
Rye Soluble 3.4 0.9 - 0.1 0.1 0.1 4.6
Insoluble 5.5 1.1 1.5 0.2 0.2 0.1 8.6
Corn Soluble 0.1 - - - - - 0.1
Insoluble 5.1 - 2.0 0.2 0.6 - 8.0
1Arabinose+xylanose; 2Mannose; 3Galactose; 4Galacturonic + glucuronic acids (Source: Choct, 1997)
Figure 1: pH profile of several fungal and bacterial
xylanases (Source: AB Vista, 2013)
Grain&feed millinG technoloGy34 | July - august 2013
FEATURE

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5. variation in these cell wall components between
the main cereals used in pig and poultry diets.
ß-glucans are present in most cereals, and
particularly prevalent in barley and oats, whilst
arabinoxylans and cellulose make up the major-
ity of the cell wall NSP in corn, wheat, rye, oats
and barley grains.
Not all enzymes are equal
The challenge when it comes to comparing
single versus multi-enzyme dosing is that each
of these NSPs are not only present in different
quantities in different feed ingredients, but are
also broken down by a different enzyme type,
and can affect digestion and subsequent animal
performance in different ways. Soluble ß-glucans
may be responsible for much of the increased
digesta viscosity that reduces digestibility in barley
diets, but it is the soluble arabinoxylans that
appear to have a similar effect in wheat. Further,
as these polysaccharides are closely associated in
the cell wall structure, anything that affects the
structure of one is likely to influence that of the
others.
Cell wall NSPs are also known to decrease
the availability of intracellular starch for break-
down within the animal gut – hence the
improvement in starch digestibility achieved by
appropriate NSP enzymes. On top of this are
the less clearly defined benefits that come from
improved populations of beneficial gut micro-
flora when incorporating certain NSP enzymes,
which release oligosaccharides, in the diet.
Very clearly, any comparison therefore needs
to take into account not just the types of
enzymes involved, but also the impact different
feed ingredients might have on the results. In
addition, the effect of any products of enzyme
activity need to be considered, some of which
(such as specific oligosaccharides) may be ben-
eficial, possibly pre-biotic, while others (such as
free sugars) may be detrimental.
Finally, keep in mind that commercial enzymes
are not pure, so a product labelled as a xylanase will
also invariably contain ß-glucanase and a number of
other activities. Any variation in dose rate between
the commercial products being evaluated must also
be accounted for, and the testing procedure able to
cope with any differing characteristics exhibited by
enzymes even of the same type.
Figure 1, for example, shows the pH profiles
for several fungal and bacterial xylanases, indicat-
ing how activity levels vary as pH changes. The
problem for any comparative testing that involves
more than one enzyme is that activity which
might appear to be similar or additive when
tested at pH5.5 may well produce completely
different results under the variable conditions
present in the digestive tract (more acid in the
stomach or gizzard, more alkaline in the small
intestine).
Do enzyme characteristics matter?
It may be argued that as long as animal
performance is improved in trials, awareness of
these differences in enzyme characteristics and
identifying exactly which activities are causal is
unimportant. However, evidence available to
date suggests that in many such comparisons,
multi-enzyme dosing fails to outperform the best
of the single enzyme products.
Even where an improvement has been
achieved, it is often not possible to determine
whether the result was due to the additional
enzyme types or, for example, more of the
original enzyme type. If a study compares
product A (a xylanase) with product B (a
multi-enzyme product containing a different
xylanase), any performance improvement from
product B may not come from the additional
enzymes, but could instead be related to other
factors, such as the
xylanase being sup-
plied at a higher
dose rate, having
improved activity, or
being more appro-
priate to the test
diet being used.
Figure 2 shows
the results of an AB
Vista broiler trial car-
ried out to investigate
the difference in per-
formance between
four commercial xyla-
nase-based products.
In this case, a single
xylanase product
(Econase XT) out-
performed both the other single xylanase prod-
uct and the two multi-enzyme products tested. It
is clear that in this trial, choosing a multi-enzyme
product is not necessarily beneficial.
In fact, what the results show is that the
characteristics and dose of the enzyme being
used are far more important than the number of
enzymes present in each product. This is critical
when it comes to choosing between products in
a commercial situation.
Furthermore, if there is no uniformity in per-
formance response to even the same enzyme
from different origins due to variation in enzyme
characteristics, then valid direct comparisons
become difficult to achieve. Table 2 lists the
minimum number of trial treatments needed
to provide a complete comparative dataset for
an example product containing three enzymes.
However, even this relies on each xylanase
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Figure 2: Comparison of bird performance (0-42 days)
when fed a range of xylanase-based enzyme products
(Source: AB Vista, unpublished)
Grain&feed millinG technoloGy36 | July - august 2013
FEATURE

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7. being from the same origin, at the same
dose rate and supplied in the same form,
with similar criteria for the ß-glucanase and
protease. Achieving such a comparison using
only commercially available products is sim-
ply not possible in the vast majority of cases.
It is also been proposed that since the
response to additional enzyme inclusion
is dependent upon the remaining fibrous
content of the diet, any such response
will be reduced with every extra enzyme
included. In the example outlined in Table
2, the ability of a ß-glucanase to poten-
tially improve performance when added
to a xylanase (treatment 5) or a protease
(treatment 7) may be irrelevant to its
effect as part of the test product – the
combination of xylanase plus protease
might simply be so effective that there is
little performance response available from
addition of the ß-glucanase.
Interestingly, in the limited number of
studies in which a more complete com-
parison has been made, it is not usu-
ally the enzyme product with the greatest
number of activities that results in the
best performance. In one 2012 study to
evaluate the effects of a xylanase and a
protease in broilers fed wheat-soy-based
diets, both enzymes improved performance
individually. However, no further improve-
ment was seen in any growth parameters
(bodyweight, feed intake or feed conver-
sion ratio) when these two
activities were combined
(Kalmendal and Tauson,
2012).
Similar results have
been seen for the inter-
actions between xylanase
and ß-glucanase in maize-
soy-based diets in broilers.
Despite the enzymes also
being tested at different
dose rates, it was found
that whilst the xylanase and
ß-glucanase alone improved
feed conversion, combining
both together resulted in
no further improvements (Cowieson et al.,
2010).
Achieving consistent results
The one situation where more consist-
ent results appear to be achievable is when
a phytase is supplemented with an NSP
enzyme, perhaps due to the greater dif-
ferences in mode of action and substrate.
Evaluation of the effects of dietary enzymes
on performance of broilers fed a maize-soy-
based diet, for example, found that the only
combination of enzymes producing a further
improvement in performance over single
enzymes was phytase plus xylanase (Walk
et al., 2011).
For the feed manufacturer, this general
lack of clarity is far from helpful, and the
comparison of single versus multi-enzyme
dosing is an area that would appear to justify
further study. In the meantime, an aware-
ness of those underlying factors which can
influence animal performance is vital when
unravelling the data that is used to pro-
mote enzyme products in the marketplace.
Remember that good data still needs correct
interpretation if the right conclusions are to
be drawn!
More InforMatIon:
Website: www.abvista.com
Table 2: Hypothetical experimental treatment design for multi-
enzyme evaluation
Treatment Enzyme activity
1
Test product - known to contain xylanase,
ß-glucanase and protease activity
2 Xylanase
3 Glucanase
4 Protease
5 Xylanase and ß-glucanase
6 Xylanase and protease
7 ß-glucanase and protease
Control No enzyme
Grain&feed millinG technoloGy July - august 2013 | 37
6th
Protein Summit 2013
Platform for Future supply, Health &Technology
For information on partnerships & exhibiting, contact: Gerard Klein Essink
gkleinessink@bridge2food.com | ph: +31 30 225 2060 | www.bridge2food.com
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International Speakers from Unilever, FAO, Rabobank, Cosucra,TNO,
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FEATURE
Kerry Agribusiness
gets retrofit bag
former
K
erry Agribusiness Feed
Mill in Farranfore,
Co. Kerry, Ireland has
recently installed a retrofit bag
former with help from Premier
Tech Chronos, UK. The retrofit
project replaces a bag forming
until that was originally supplied
more than 20 years ago.
Kerry Agribusiness provides a
range of products and services
that are aimed at optimising
profitability at a farm level.
From their Farranfore Feed
Mill Kerry Agribusiness has
been supplying top quality
animal feeds across the Munster
region.
As part of the retrofit project,
Premier Tech Chronos
service engineers supported
the Kerr y Agribusiness
engineers who re-installed and
re-commissioned the SSV Bag
Former. One of the major aims
of the retrofit project was to
ensure continued optimum
packing outputs were achieved
on the previously supplied bag
placing and bag preparation
system, which handles a diverse
range of animal feed products.
Since early 2003, genuine spare
parts matched to the original
Chronos Richardson design
authority documentation and
drawings have been supplied by
fellow Premier Tech company;
Premier Tech Chronos. That
experience and expertise is
still available today for service
and aftermarket support for the
entire company product range.
This expertise and capability
was an important factor in the
company’s ability to support
Kerry Agribusiness on this
particular project.
Bayer launches
product guide
N
ow is prime time for
growers to plan their
pre-harvest grain storage
treatments and Bayer is making
this process easier by the launch of
a new grain protectant guidebook.
The manual provides farmers and
pest controllers with all they need
to know about Bayer’s K-Obiol®
insecticide.
“Storinggraincomeswithavariety
of challenges,” says Ken Black,
national account manager for rural
hygiene, Bayer.
“There are three factors that
influence the quality of the grain:
temperature, moisture content
and storage period. The greater
the quantity of grain, the greater
the risk of infestation.”
Anestimated90percentoffarmgrain
storesharbouratleastonespeciesof
insectknowntoinfestgrain.“When
grain prices are high and with the
possibility of growers choosing to
storetheirgrainforlongeraplanned
approach to good grain storage is
essential.Wehopeourguidewillhelp
growersmakethebestdecisionsfor
protectingtheiryield.”
To order a K-Obiol grain
protectant guide book
email pestcontrolexpert@
bayercropscience.com
37% of global overall diets are
made up of animal products in North
America and Western Europe
5-7% of global overall diets are
made up of animal products in Sub-
Saharan Africa and South Asia
38% of global cereal is used for
animal feed
25 units of feedstuff are used to
produce one unit of livestock output
34.8 million tonnes of fish was used
for terrestrial livestock feed in 2004
7times more feed is demanded globally
for ruminants than monogastrics
2 crop categories dominate global
livestock feed: cereals and oil crops.
Source: The Impact of Industrial Grain
Fed Livestock Production on Food
Security: an extended literature review,
Alpen-Adria University, Austria
NUMBERCRUNCHING
Animal feed
NewsJuly - August 2013NEWS
Grain&feed millinG technoloGy8 | July - august 2013
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Ann A5 victam 0212.indd 1 21/02/12 15:37:04

8. www.gfmt.co.uk
LINKS
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A subscription magazine for the global flour & feed milling industries - first published in 1891
INCORPORATING PORTS, DISTRIBUTION AND FORMULATION
In this issue:
• Pig feed
pelletizing
technology
• Feed focus
Cattle
• Exploring
the challenge
of single
versus multi-
enzyme dosing
comparisons
July-August2013
• Improving
poultry health
and production
efficiency with
probiotics
• Aflatoxins in
Europe:
a new risk in maize
production?
• Sweeping
changes
to OSHA’s
sweep auger
enforcement
first published in 1891
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