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Exploring the challenge of single versus multi-enzyme dosing comparisons

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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.

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Exploring the challenge of single versus multi-enzyme dosing comparisons

  1. 1. Digital Re-print - July | August 2013 Exploring the challenge of single versus multi- enzyme dosing comparisons Grain & Feed MillingTechnology is published six times a year by Perendale Publishers Ltd of the United Kingdom. All data is published in good faith, based on information received, and while every care is taken to prevent inaccuracies, the publishers accept no liability for any errors or omissions or for the consequences of action taken on the basis of information published. ©Copyright 2013 Perendale Publishers Ltd.All rights reserved.No part of this publication may be reproduced in any form or by any means without prior permission of the copyright owner. Printed by Perendale Publishers Ltd. ISSN: 1466-3872
  2. 2. 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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  4. 4. 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 FROM IAOM’S 2013 ANNUAL CONFERENCE THROUGH THE IAOM LIVE LEARNING CENTER EXPERIENCE EDUCATIONAL SESSION RECORDINGS Recordings are complimentary for all 2013 Annual Conference attendees as well as IAOM members. 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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  6. 6. 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: 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 | ph: +31 30 225 2060 | 24 & 25 September 2013, Rotterdam International Speakers from Unilever, FAO, Rabobank, Cosucra,TNO, Rousselot, Innova Market Insights, Koch Membrane Systems, Bühler, Tereos Syral,Wageningen University,True Price and many more... 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@ 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 VIGAN Engineering s.a. • Rue de l’Industrie, 16 • B-1400 Nivelles (Belgium) Phone : +32 67 89 50 41 • Fax : +32 67 89 50 60 • Web : • E-mail : VIGAN manufactures dry agribulk materials handling systems: • Portable pneumatic conveyors or grain pumps (100 - 250 tph); • Pneumatic Continuous barge & Ship Unloaders (160 - 800 tph); • Mechanical Continuous Ship Unloaders (up to 1,500 tph); • Mechanical loaders (up to 1,200 tph). as well as complete storage systems in ports and the agricultural industries. From project design to complete turnkey bulk handling solutions and port terminals with mechanical and/or pneumatic reliable and cost effective equipment. 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  7. 7. LINKS • See the full issue • Visit the GFMT website • Contact the GFMT Team • Subscribe to GFMT 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 This digital Re-print is part of the July | August 2013 edition of Grain & Feed Milling Technology magazine. Content from the magazine is available to view free-of-charge, both as a full online magazine on our website, and as an archive of individual features on the docstoc website. Please click here to view our other publications on To purchase a paper copy of the magazine, or to subscribe to the paper edi- tion please contact our Circulation and Subscriptions Manager on the link adove. INFORMATION FOR ADVERTISERS - CLICK HERE Article reprints All Grain & Feed Milling Tecchnology feature articles can be re-printed as a 4 or 8 page booklets (these have been used as point of sale materials, promotional materials for shows and exhibitions etc). If you are interested in getting this article re-printed please contact the GFMT team for more informa- tion on - Tel: +44 1242 267707 - Email: or visit