Phytases beyond phosphate, how inositol improves feed conversion ratio in poultry

Lead Research Organisation: University of East Anglia
Department Name: Biological Sciences

Abstract

We feed large amounts of cereal grains and beans to animals. The efficiency with which animals convert feed to body weight is the most important measure of the productivity of intensive animal agriculture. Feed conversion ratio (FCR) is the amount of feed (kg) required to increase body mass by 1kg. Of the animals, fish, poultry, pigs and cattle that are farmed intensively in the UK, fish are the most efficient converters of feed into body mass, with a FCR of 1. Poultry, depending on the species and feeding period, have a FCR of < 2, while pigs and cattle are much less efficient.

One way in which farmers can improve FCR is by the addition of enzymes to animal feed. Our partner in this project, AB Vista Feed Ingredients, a division of AB Agri, a part of Associated British Foods, ABF, is involved in the development and distribution of feed enzymes including phytases. Phytases are essential for efficient use by the animal of a phosphate-rich molecule that is the major storage form of phosphorus in feed components of plant origin like grains and soy beans. The phosphate rich molecule, phytate, or inositol hexakisphosphate, contains six phosphates for every inositol. Inositol has some similarity to sugars, but, unlike sugars, we know little of its digestive use to the animal. In contrast, phosphate is known to be hugely important to mineral nutrition, bone development and growth of animals.

Companies in ABF including AB Vista Feed Ingredients have substantial programs of research for improvement of phytases in respect of their application and their intrinsic properties. There are several drivers of this research: improvements in animal growth afforded by access to phytate-bound phosphate in the dietary feedstuff and avoidance of anti-nutrient effects of dietary phytate. The use of enzymes to access phosphate in feeds means that less of this essential nutrient needs to be added as rock phosphate (commonly dicalcium phosphate). It has been estimated that the addition of less than $1 worth of enzyme spares rock phosphate costs of $4-6/tonne of feed.

Many companies undertake poultry feeding trials to show that their enzymes work. A common approach is to feed animals with different amounts of phytase in a randomized trial design and to measure animal performance. While many trials highlight the value of phytase addition to animal feed, few companies measure phytate degradation by analysing gut contents (digesta) for phytate and its inositol phosphate degradation products. AB Vista Feed Ingredients does because the approach explains how their products work, enabling development of better products.

New research suggests that inositol released from feed during digestion is beneficial to animal growth. The mechanisms by which inositol improves animal growth is unclear. While we may reasonably assume that inositol generated during digestion enters the blood and is transported to tissues very few scientific studies have been made of the subject.

The applicants, Charles Brearley, Gabriel Mutungi and AB Vista Feed Ingredients have over 20y experience, respectively, of inositol phosphate and inositol analysis, of muscle physiology including of farmed poultry, and of the use of feed enzymes in animal nutrition. Our programme of research is designed to reveal how inositol, released by phytases, improves nutrition in poultry. We will exploit newly developed methods that allow measurement of inositol in feed, digesta and plasma and adapt these methods to analyse inositol in muscle tissue. Additionally by analysis of the physiology of muscle, we expect to explain how inositol improves muscle growth, identify the key benefits of phytases and so improve the phytase dosing regimens of farmed poultry with consequence for the environment and the consumer.

Technical Summary

Our published work shows correlation between exogenous enzyme- (phytase) mediated release of inositol from feed, measured in gizzard contents, and body weight gain in poultry. To provide a deeper understanding of the effects of inositol and its practical benefits to animal nutrition, we will:
1) Measure inositol content across the GI tract, in gizzard and ileum contents as a function of phytase dose. Samples taken from the gizzard and ileum, provided by the industrial partner from feeding trials designed for this project, will be analysed by 2d-HPLC with pulsed amperometric (PAD) detection; 2) Measure inositol content of plasma as a function of phytase dose. Plasma provided from the above feed trials will be analysed by 2d-HPLC-PAD; 3) Measure inositol content of muscle tissue as a function of phytase dose. Muscle tissue provided by our industrial partner from feed trials will analysed by 2d-HPLC-PAD; 4) Measure inositol phosphates across the GI tract, gizzard and ileum, in plasma and in muscle tissue. We will apply a range of HPLC separation techniques either with suppressed ion conductivity detection or with post-column complexation of inositol phosphates to the feed and digesta samples provided by our industrial partner; 5) Correlate measurements 1, 2, 3, and 4 with indices of animal performance. The parameters of animal performance that we will measure include, feed conversion ratio, body weight gain, tibia strength and/or mineralisation; 6) Undertake assessment of muscle physiology as a function of phytase dose, correlating it with circulating (plasma) and tissue inositol content. We will measure: slow vs fast twitch bundle composition; fat, carbohydrate and protein levels; inositol content; inositol phosphate content and Akt/mTOR pathway activation.

Planned Impact

Supplementation of animal feed with enzymes improves animal nutrition and this explains why approximately 90% of the pig and poultry feeds manufactured worldwide include exogenous enzymes. The feed enzyme market was worth $650m in 2010 with annual growth in the past of 13% (2010, Barletta in Bedford and Partridge: Enzymes in farm Animal nutrition). The internal estimates of our industry partner suggest that growth might even have increased. Of the principal enzyme groups in the feed enzyme market, non-starch polysaccharases and phytases, the latter account for approximately 50% of the sales volume.

Our proposal is a collaborative program of research between UK industry, AB Vista Feed ingredients, a division of the AB Agri group of Associated British Foods, and academia. It involves fundamental and applied research in the area of sustainable agriculture.

The research described in this proposal affords opportunity for a radical reappraisal of the physiological explanation of the efficacy of phytase adjuncts to animal feed. Our proposal addresses a research problem that the feed industry and the academic partners have identified and our proposal offers solution to that problem. Put simply, with ever increasing pressure on food and animal feedstuffs, feed conversion ratio and/or body weight gain is/are the most critical index of the efficiency of intensive animal agriculture. This proposal goes to the heart of feed conversion ratio and the contribution that enzyme adjuncts make to that. BBSRC has identified sustainable agriculture as a high-level priority area in its Strategic Plan 2010-2015.

The potential impacts of this work are multifaceted. Because we will publish our research in poultry nutrition journals, read by the whole industry, from farmer to feed mill to integrated poultry producers to animal nutrition scientists in academia and the regulatory bodies, our work will inform all companies who undertake their own feed trials in poultry and will enhance the research capacity, knowledge and skills of businesses developing products in this field.

Our industrial partners attend several applied animal nutrition conferences worldwide each year and regularly chair sessions or give plenary talks thereat. Consequently, our research will be prominently reported to experts in the field.

AB Vista has sophisticated marketing strategies, dedicated marketing specialists and a global reach and reputation that ensures the outputs of its research are disseminated widely to, and by, feed industry analysts whose own outputs are keenly followed by the industry and also by commodity traders.

Advertisement of the outputs of our research will also include direct press releases by AB Vista Feed Ingredients.

The impacts to be expected extend to improvements in animal nutrition and reductions in cost in the food supply chain, amelioration of phosphate outputs to the environment and reductions in mined mineral inputs to agriculture. The impacts are acute in the context of intensive poultry production. Because phytases are employed world-wide, this work has global relevance.

This project will provide excellent training opportunities for the PDRA in analytical biochemistry and animal nutrition and will give the scientist unrivalled access to expertise in the commercial arena of a biotechnology industry with agricultural focus. The Norwich Research Park (NRP) is an exemplar of excellent plant research and the postdoctoral scientist will have ample opportunity to extend his/her presentational skills and network within the local scientific community. The collaboration between the UEA groups and the industrial partners will extend these opportunities by contributing towards the pool of highly skilled researchers who can contribute with a skill set demanded by the animal feed industry to the UK economy. This project enhances the reputation of the NRP as an industry-aware research environment.

Publications

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Neal AL (2017) Land-use influences phosphatase gene microdiversity in soils. in Environmental microbiology

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White G (2020) Regioisomeric Family of Novel Fluorescent Substrates for SHIP2. in ACS medicinal chemistry letters

 
Title Feeding Animals Sustainably 
Description An infographic comparing the feed efficiency (Feed Conversion Ratio) of different animal production systems that do or do not use phytase adjuncts 
Type Of Art Artefact (including digital) 
Year Produced 2016 
Impact not known 
URL https://www.uea.ac.uk/research/explore-uea-research/feeding-animals-sustainably
 
Description With our industrial partnters, we have established that phytases added to the feed of non-ruminant (monogastric) animals, poultry and pigs, increase the growth performance of these animals. We show that the performance of these animals is related to the digestion of a major form of phosphate, phytate, present in the plant-based feed of these animals.

We show that some of the increases in growth performance are associated with the use of phosphate released from phytate.

We further show that some of the increases in growth performance are associated with the release of a molecule called inositol into the gut.

We developed methods to measure phytate digestion by the animals.

We developed methods to measure the inositol released during digestion of phytate, in feed and in animal digesta.

We further developed methods to measure inositol in blood (plasma) and in tissues such as liver, muscle, kidney, showing that phytase addition to a animal diets increases inositol in some of these tissues.

Our work provides a mechanistic explanation of the action of phytases added to animal diets, ie. how they improve animal nutrition.
Exploitation Route The methods we have developed for measurement of inositol and inositol phosphate digestion products of phytate in food, digesta, plasma and tissues, are relevant to biomedical disciplines, particularly the role of inositol in human health.
Sectors Agriculture, Food and Drink,Environment

URL https://www.uea.ac.uk/research/explore-uea-research/feeding-animals-sustainably
 
Description The research and development of our industrial partner is influenced by the findings of our collaborative research, this impacts the strategy and economic success of our partner. The research direction of our partner takes direct and responsive account of our findings.
Sector Agriculture, Food and Drink
Impact Types Economic

 
Description BBSRC NRP iCASE DTP
Amount £89,000 (GBP)
Funding ID A Mechanistic Explanation of the Benefits of Phytase to Sustainable Agriculture - BREARLEY 
Organisation Biotechnology and Biological Sciences Research Council (BBSRC) 
Sector Public
Country United Kingdom
Start 10/2018 
End 09/2022
 
Description Direct Funding from Industrial Partner
Amount £105,410 (GBP)
Funding ID UEA RENU00529 
Organisation AB Vista 
Sector Private
Country United Kingdom
Start 09/2016 
End 06/2017
 
Description Direct Industrial Funding
Amount £161,195 (GBP)
Funding ID AB Vista: UEA RENU00529 - Direct Funding from Industrial Partner 
Organisation AB Vista 
Sector Private
Country United Kingdom
Start 06/2017 
End 12/2018
 
Description Industrial Research AB Vista
Amount £147,435 (GBP)
Funding ID UEA RENU00529 
Organisation AB Vista 
Sector Private
Country United Kingdom
Start 11/2016 
End 11/2017
 
Description NERC ENV EAST CASE DTP
Amount £89,000 (GBP)
Funding ID The biodiversity of organic phosphate (phytate) cycling in soils 
Organisation University of East Anglia 
Department EnvEast NERC Doctoral Training Partnership
Sector Academic/University
Country United Kingdom
Start 10/2017 
End 09/2021
 
Description INDUSTRIAL CASE AWARDS 
Organisation AB Vista
Country United Kingdom 
Sector Private 
PI Contribution We have engaged our industrial partners as CASE partners to NERC ENV EAST DTP and BBSRC NRP DTP studentships started Oct 2017 and Oct 2018, respectively. We have further engaged other companies (Drayton Animal Health Ltd) in our collaborative work with AB Vista. Drayton Animal Health Ltd is a partner/sub-contractor to the BBSRC NRP iCASE DTP studentship. Our PhD students and Post Docs have given talks to our industrial partners at a 'talent-spotting' conference that AB Vista hold. Our PhD students and Post Docs have also attended forward planning/collaboration meetings with Research Directors of AB Vista, AB Enzymes (Germany) and Roal Oy (Finland) March 2017 (Gerrard's Cross, UK) and Feb 2018 (Heathrow, UK). Our industrial partners have flown in from Germany and Finland to these meetings. Two PhDs funded by AB Vista have been succesfully examined in Jan 2017 and Jan 2018, one awarded July 2017, the other awarded - July 2018. Another PhD student funded wholly by AB Vista will be examined March 2019. Dr Brearley was an industry paid delegate to the Third Phytase Summit, Nov 2016, Miami, USA. AB Vista are additionally supporting as iCASE partner another application (Complete Dephosphorylation of Phytic Acid: Identification of Histidine Acid Phosphatases with Novel Phytase Activity for Use in Animal Feeds) to the last round (March, 2019) of the current BBSRC NRP iCASE DTP programme.
Collaborator Contribution In-kind and direct cash contributions to RCUK studentships of > £130000.
Impact The ENV EAST NERC CASE DTP studentship detailed above engages soil scientists and environmental microbiologiest at Rothamsted Research (BBSRC) in a phytase discovery project http://www.enveast.ac.uk/
Start Year 2017
 
Description INDUSTRIAL RESEARCH/CONSULTANCY 
Organisation AB Vista
Country United Kingdom 
Sector Private 
PI Contribution Development and refinement of methods for analysis of inositol phosphates, inositol and saccharides in animal feed, digesta, plasma and tissues. Provision of analytical data for verification of the efficacy of enzyme adjuncts (phytases and xylanases) in feed trials. Contribution of conference manuscripts and other publications with our industrial partner.
Collaborator Contribution Commission of animal feed trials in UK, Europe, ASIA and USA. Import of materials to UK for analysis at UEA.
Impact Multiple outputs in the form of conference papers and research papers (submitted - or accepted). All either co-authored with our industrial partners and other international academic partners, or with international authors not connected to AB Vista. Industrial partner authors include Carrie Walk, Mike Bedford, Imke Kühn. UEA authors include Charles Brearley, Hayley Whitfield, Michael Smith H. Lu, I. Kühn, M. Bedford, H. Whitfield, C. Brearley, O. Adeola, and K. M. Ajuwon (2019) Effect of phytase on intestinal phytate breakdown, plasma inositol concentrations and glucose transporter type 4 abundance in muscle membranes of weanling pigs (JAS-2019-3356) Journal of Animal Science. Submitted Feb 2019 Attributed to BBSRC LINK BB/N002024/1 Phytase superdosing increased yolk mineral concentration while decreasing yolk inositol concentration from breeder hens aged 35 or 40 weeks C. A. Granghelli1, C. L. Walk2, L. F. Araujo1, S. M. Silva1, M. L. Cuadros1, Y. G. A. Sartore1, M. T. Dias1, C. Brearley3, M. Smith3, and C. S. S. Araujo1 1 University of Sao Paulo, Pirassununga, Brazil 2 AB Vista, Marlborough, Wiltshire UK SN8 4AN 3 University of East Anglia, Norwich, UK European Symposium on Poultry Nutrition, Poland, 2019. Phytase dose fed to breeder hens has an influence on yolk inositol concentration, chick quality and hatchability, and early chick growth rate C. L. Walk1, C. A. Granghelli2, C. S. S. Araujo2, L. F. Araujo2, F. A. Roque2, B. G. S. Leite2, C. Brearley3, and P. H. Pelissari2 1 AB Vista, Marlborough, Wiltshire UK SN8 4AN 2 University of Sao Paulo, Pirassununga, Brazil 3 University of East Anglia, Norwich, UK European Symposium on Poultry Nutrition, Poland, 2019. Claus Krogh Madsen, Charles Alistair Brearley & Henrik Brinch-Pedersen (2019) Lab-scale preparation and QC of phytase assay substrate from rice bran. Analytical Biochemistry: Methods in Biological Sciences submitted Attributed to BBSRC LINK BB/N002024/1 Laird S, Kühn I, Bedford MR, Whitfield H, Miller HM, Sampling duration and freezing temperature influence the analysed gastric inositol phosphate composition of pigs fed diets with different levels of phytase, Animal Nutrition Journal, https://doi.org/10.1016/j.aninu.2018.12.003. Attributed to AB Vista
Start Year 2017
 
Description Display at Royal Norfolk Show 29-30 June 2016 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Public/other audiences
Results and Impact All members of the laboratory attended the Innovation Hub at the Royal Norfolk with a display and advertising material highlighting our collaborative research in animal nutrition. Our display named our industrial partner (AB Vista Feed Ingredients Ltd) with whom all publicity material was agreed.
Year(s) Of Engagement Activity 2016
 
Description Industry organized conference 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Industry/Business
Results and Impact Paid attendee at Business organized conference 3rd International Phytase Summit, Nov 2016, Miami
Year(s) Of Engagement Activity 2016
URL https://www.abvista.com/Innovation/International/IPS3.aspx
 
Description Norwich Science Festival at The Forum, Norwich 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Public/other audiences
Results and Impact Posters and laboratory-type demonstrations were made to public and school audiences at the Norwich Science Festival (24/10/16).
Year(s) Of Engagement Activity 2016
 
Description University Open Day 
Form Of Engagement Activity Participation in an open day or visit at my research institution
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Schools
Results and Impact Laboratory demonstrations were provided and Poster presentations were made to prospective students and their parents/guardians/company on the subject of our industrial-sponsored research at University Open Days
Year(s) Of Engagement Activity 2016