Digital Re-print July | August 2013

Exploring the challenge of single versus multienzyme dosing comparisons

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FEATURE

Exploring the challenge of single versus multi-enzyme dosing comparisons

by Dr Helen Masey ONeill, research manager, and Tiago dos Santos, global technical manager, AB Vista, United Kingdom
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.

Table 1: Types and estimated levels of the main fibre polysaccharide components present in key cereal grains % of dry matter Cereal Xylan1 -Glucan Cellulose Mannan2 Galactan3 Uronic acids4 0.2 0.2 0.1 0.7 0.1 0.1 Total

Wheat Barley Oats Rye Corn

Soluble Insoluble Soluble Insoluble Soluble Insoluble Soluble Insoluble Soluble Insoluble

1.8 6.3 0.8 7.1 1.0 9.2 3.4 5.5 0.1 5.1

0.4 0.4 3.6 0.7 2.8 0.5 0.9 1.1 -

2.0 3.9 8.2 1.5 2.0

0.2 0.2 0.1 0.1 0.2 0.2

0.2 0.1 0.1 0.1 0.2 0.4 0.1 0.2 0.6

2.4 9.0 4.5 12.2 4.3 19.1 4.6 8.6 0.1 8.0

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 components in the diet. This fibrous content will differ depending on the feed ingredients used, however, with the main constituents being cellulose, arabinoxylans, mixed-linked -glucans, glucomannans, galactomannans and arabinans. The values in Table 1 illustrate the
34 | July - august 2013

1Arabinose+xylanose; 2Mannose; 3Galactose; 4Galacturonic + glucuronic acids (Source: Choct, 1997)

Figure 1: pH profile of several fungal and bacterial xylanases (Source: AB Vista, 2013)

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FEATURE variation in these cell wall components between the main cereals used in pig and poultry diets. -glucans are present in most cereals, and particularly prevalent in barley and oats, whilst arabinoxylans and cellulose make up the majority of the cell wall NSP in corn, wheat, rye, oats and barley grains. Figure 1, for example, shows the pH profiles for several fungal and bacterial xylanases, indicating 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).

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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 -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 breakdown 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-

Do enzyme characteristics matter?

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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 supplied at a higher dose rate, having improved activity, or being more appropriate to the test diet being used. Figure 2 shows the results of an AB Vista broiler trial carried out to investigate the difference in performance between four commercial xylaFigure 2: Comparison of bird performance (0-42 days) nase-based products. when fed a range of xylanase-based enzyme products In this case, a single (Source: AB Vista, unpublished) xylanase product (Econase XT) outflora when incorporating certain NSP enzymes, performed both the other single xylanase product and the two multi-enzyme products tested. It which release oligosaccharides, in the diet. Very clearly, any comparison therefore needs is clear that in this trial, choosing a multi-enzyme to take into account not just the types of product is not necessarily beneficial. In fact, what the results show is that the enzymes involved, but also the impact different feed ingredients might have on the results. In characteristics and dose of the enzyme being addition, the effect of any products of enzyme used are far more important than the number of activity need to be considered, some of which enzymes present in each product. This is critical (such as specific oligosaccharides) may be ben- when it comes to choosing between products in eficial, possibly pre-biotic, while others (such as a commercial situation. Furthermore, if there is no uniformity in perfree sugars) may be detrimental. Finally, keep in mind that commercial enzymes formance response to even the same enzyme are not pure, so a product labelled as a xylanase will from different origins due to variation in enzyme also invariably contain -glucanase and a number of characteristics, then valid direct comparisons other activities. Any variation in dose rate between become difficult to achieve. Table 2 lists the the commercial products being evaluated must also minimum number of trial treatments needed be accounted for, and the testing procedure able to to provide a complete comparative dataset for cope with any differing characteristics exhibited by an example product containing three enzymes. However, even this relies on each xylanase enzymes even of the same type.
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Kerry Agribusiness when thesesystem, two which a diverse profitability at ratio) a farm level. being from the same origin, at the same sion Table handles 2: Hypothetical experimental treatment design for multiwere combined range of animal feed products. From their activities Farranfore Feed dose rate and supplied in the same form, NUMBER enzyme evaluation gets retrofit CRUNCHING and Since early 2003, genuine spare Mill Kerry Agribusiness has Tauson, with similar criteria bag for the -glucanase and (Kalmendal Treatment Enzyme activity 2012). parts matched to the original been supplying top quality protease. Achieving such a comparison using former Animal feed

Similar results Chronos have Richardson design animal feeds across the Munster only commercially available products is simTest product - known to contain xylanase, authority documentation and ply noterry possible in the vastFeed majorityregion. of cases. been seen for the interAgribusiness 37% of global overall diets are 1 -glucanase and protease activity actions between drawings have been supplied by As part of the retrofit project,xylanase It isM also that the ill been i n F proposed ar r a n fore , since made up of animal products in North 2 Xylanase -glucanase fellow Premier Tech company; r e m i e r and Te c h C h r o n oin s maizeresponse additional enzyme inclusion Co.to Kerry, Ireland has P America and Western Europe 3 Glucanase soy-based diets in broilers. Premier Tech Chronos. That supported is dependent upon the remaining fibrousengineers recently installed a retrofit bag service Protease is Despite the also and4 expertise t h e Ke r r y Agrib u s i enzymes n e s s experience content of help the from diet, Premier any such response former with 5-7% of global overall diets are 5 Xylanase and -glucanase being tested at still available today for service re-installed anddifferent will be reduced with extraengineers enzyme who Tech Chronos, UK. Theevery retrofit made up of animal products in Subit Bag was found and aftermarket support for the the SSV 6 Xylanase and protease included. In the example outlinedre-commissioned in Table dose rates, project replaces a bag forming Saharan Africa and South Asia that whilst the aims xylanase and company product entire range. Former. of the major 2, the ability of a -glucanase poten-One until that was originally supplied to 7 -glucanase and protease -glucanase improved This expertise and No capability of added the retrofit project alone was to tially than improve performance when more 20 years ago. 38% of global cereal is used for Control enzyme feed conversion, combining was an important factor in the optimum to a xylanase (treatment 5) or a ensure proteasecontinued animal feed bothwere together resulted in companys ability to support outputs achieved (treatment 7) mayprovides be irrelevant to its Kerry Agribusiness a packing no further improvements (Cowieson et al., on Agribusiness on the previously supplied bag Kerry effect of asproducts part of and the services test product the lackthis of clarity25 is far from helpful, and range units of feedstuff arethe used to bag preparation particular project. combination of atxylanase plus placing proteaseand2010). comparison of single one versus multi-enzyme that are aimed optimising produce unit of livestock output might simply be so effective that there is dosing is an area that would appear to justify little performance response available from Achieving consistent results further study.34.8 In the million meantime, awaretonnesan of fish was used stores harbour at least one species of thosefor addition of the -glucanase. ness of underlying which can to know about The Bayers K-Obiol terrestrial factors livestock feed in 2004 one situation where more consistknown to grain. When animal performance is vital when Interestingly, in the limited number of ent results appear to beinsect influence insecticide. achievable is infest when grain prices and unravelling with the studies in which a more complete com-grain the data more that feed is used to proStoring with a comes phytase isa variety supplemented with are an high NSP 7 times is demanded globally of growers parison has been made, it is not usu- enzyme, mote to enzymefor products in than the monogastrics marketplace. of challenges, says Ken Black, perhaps due possibility to the greater dif- choosing ruminants store their for longer a planned ally the ow enzyme product with the national greatestaccount Remember that good data still needs correct manager for rural ferences in mode of action and grain substrate. is prime time for to enzymes good grain storage is number growers of activities thattheir resultshygiene, in theBayer. interpretation if the right conclusions are toglobal Evaluation of the effectsapproach of dietary to plan 2 crop categories dominate essential. We hope our guide will help best performance. In one 2012 study to areon be drawn! There three factors that performance of broilers fed a maize-soypre-harvest grain storage livestock feed: cereals and oil crops. make theonly best decisions for evaluate the effects a xylanase and a the influence quality of the grain: growers based diet, for example, found that the treatments and Bayer of is making their yield. protease in broilers fed wheat-soy-based moisture content combination of enzymesprotecting producing a further this process easier by the launch of temperature, Source: The Impact of Industrial Grain To order K-Obiol grain diets, both protectant enzymes improved performance and storageimprovement period. The greater in performance overa single a new grain guidebook. Fed Livestock Production on Food p r o xylanase t e c t a n t (Walk guide b oore o k InforMatIon individually. However, no further improvethe quantityenzymes of grain, the wasgreater phytase plus M Security: : an extended literature review, email pestcontrolexpert @ ment was seen in any growth the risk of infestation. et al., 2011). The manual provides farmers and parameters Website: www.abvista.com Alpen-Adria University, Austria bayercropscience.com (bodyweight, feed An converestimated 90 percent of farm For the feedgrain manufacturer, this general pest controllersfeed with intake all they or need

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