NZ715516B2 - Animal feeds and feed premixes containing betaine hydrochloride and a phytase. - Google Patents

Abstract

The invention relates to poultry feeds and feed premixes containing synergistically effective amounts of betaine hydrochloride (BCHl) and a phytase, provided in a ratio of about 0.5 to 5g BHCl per kg of feed : 100 to 5000 phytase unit (FTU) per kg of feed.

Description

Animal feeds and feed premixes containing betaine hydrochloride and a phytase.

Field of the ion The invention relates to formulation of animal feed, and to assessment of nutrient utilisation and growth nance, especially with regard to monogastric animals and livestcckg U1 including poulny.

Background of the invention Reference to any prior art in the specification is; not, and should not be taken an. an acknowledgment or any form of suggestion that this prior art forms part of the common general dge in Australia or any other jurisdiction or that this prior art could reasonably be l0 expected to be ascertained, underetood and regarded as relevant by a person skilled in the art stric animal diets generally e nutrients for the , reproduction and health of an animal. These nutrients may take the form of proteins and amino acids: carbohydrates, fats, minerals and vitamins. “Macronutrients” are. generally understood as meaning those components of a diet that provide. the bulk of energy and protein. for metabolism.

These components are generally proteins, carbohydrates, fats and oils and fibre. nutrients” generally provide the necessary cofactors for metabolism to occur. These- components are generally mineral 3‘ vitaminS, and amino acids. As an examplefi y diets are composed. primarily ot‘ a mixture of several foodstuffs that contain macromutritive componente, examples of which include cereal grains, n meal, animal by —pr0duc~ts (such as blood and bone), fats and micro-nutritive components; including mineral and vitamin premixes. Another micromutritive component, anhydrous hetaine has at] so been used as a source of methyl groups.

Other non nutritive components may be added to an animal diet for a variety of purposes Examples e pigment (Such as xanthophyll), growth factors, anti-microbial agents and enzymes. One enzyme having increasing usage is ph‘ytase. Phytase is used, particuiarly in pork {\J L11: and poultry farming as a feed additive for the e of releasing phosphate that is bound to dietary phytate, Some of these non nutritive components may assist. in the growth mance improvements and ncy offered utilisation.

In some countries, non-nutritive components may be combined and sold as a ore-mix formulation to a stock feed producer, who then mixes the pro—mix with nutritive components (for example , y forming a finished animal food product that is sold to a farmer or animal producer. Alternatively, the lire—mix formulation may he sold directly to a farmer or animal producer who will then mix the pie—mix with nutritive componente to form an animal feed for feeding to his stock; Sometimes these pro-mix formulations. may he referred to as “feed concentrates” or “feed additive mixes.’ The nutritive and nonmutritive components of animal. feeds are a significant input. cost in the farming of monogas’rric animals. As an example the prioee in the poultryr meat market are currently high due to high prices for feed grains.

One m is that a ion in nutritive components, which might minimise costs of one or more nutritive inputs, in the absence of appropriate adjustments to an animal diet, can deletenously impact on growth perfomtanoe. Further, while ments that reduce the amount of nt components might be overcome by provision of non nutritive components that improve efficiency of feed utilieation, these non nutritive components may eimply represent another input cost.

There is a need to adapt animal feed so as. to minimise higher input costs, including input: costs relating to nutritive and/or non nutritive eomponents, as this would increase profitability of livestock production y of the invention The ion seeks to address the above identified need, anchor to provide improvements in animal feeds, and in one embodiment provides an animal feed including: - a nutrient component including one or more of a carbohydrate, fat and protein; — a phytase; — betaine hydrochloride (herein BHCl); n BHCl and phytase are ed in a ratio of amounts- ot‘BHCl : phytase so that the apparent. metabolisahle energy (AME) value of the animal feed equals the AME value of the nutrient component. in the presence of the amounts of BHCI and phytase.

In another embodiment there is provided a process for producing an animal feed including the step of: ~ combining a nutrient component including one or mere 01‘“ a carbohydrate, fat and protein with a phytase and. BBC], wherein the BHCl and phytase are provided in a ratio of” amounts of BH‘CI : phytase 30 that the AME value of the animal feed equals the AME value of the nutrient component. in the U1 presence ef the amounts of BHCI and phytase; thereby producing the animal feed.

In another embodiment there is ed a precess for increasing the AME value of an animal feed including the stepe 0t“. prnviding an animal feed in the form 0f a nutrient ent including (me or more if 1. f) carbohydrate, fat and protein? and a phytase; ing the animal feed with EH Cl; wherein the BHCl is prm‘ided in a ratio 0f amounts OfBl-ICI : phytese so that the AME value of the animal feed equals the AME value of the nutrient enmponent in the presence of the amounts 0f BHCl and phytase.

In another embndiment there is provided an animal feed premix, said premix consisting 0f”; - a rient component; - wherein the nan-nutrient ent includes BHC‘l and phytase.

In another embodiment there is previded an animal Feed premix, said premix consisting — a non-nutrient mmpnnent; - a. utrient mmpenen‘t; ~ wherein the non—nvnttie‘nt enmpenent includes BHCl and phytase.

In another ment there is provided a process for producing an animal feed b.) L): including the Step of: —comhining a nutrient cemponent including ene er more of a carbohydrate, fat and protein with an animal feed premix. described abeve.

Further aspects of the present invention and further embodiments ofthe aspects described in the preceding paragraphs will become apparent from the following description, given by way ef‘example and. with reference to. the accompanying drawings.

Detailed ptinn {if the ments It will be tood that the invention disclosed and defined in. this specification extends to all alternative eembinatiens at two or more of the individual features mentiened or evident from the text or drawings. All of these different combinations censtitute various alternative aepects ofthe invention.

As described herein, the inventors have found a istic response in energy ation arising from the combination of betaine hydrochloride {BHCD with an animal feed including a phytase. The inventm‘s have also found that the improved energy utilisation translates tn an improved growth performance. 1.3 In more detail, the inventors have found that the amount of metabolisahle energy in an animal feed can be. increased by providing istically effective s of BHCi. and phytase to an animal feed. ‘Memi‘mltrnble energy is simply that component of tntal energy of an animal feed that is metabnlised by an . Energy that is net metabnlisahle is “excreted energy“ Another way of defining brn’isablra energy” is to refer to an “apparent fi‘lé‘fflbt'JZitEabZG energy 'vrtlzte’ or ‘AME traine’. An ‘AME value’ is simply the gross energy of the feed consumed minus the gross energy ned in the faeces, urine; and gaseous products of digestion.

As described herein, the ors have shown that the energy of an animal feed containing synergistic amounts of BHCl and phytase is greater than the Sum of the energy utilisation in a feed containing BHCl only} or phytase only. In this content, the inventors have found a synergistic relaticnship as between BHCl and phytase in the context of energy utilisatien. Accordingly; the AME. value of an animal feed containing the BHlephytase cnmbinatinn is greater than the AME value cf a. feed which tines net contain this combinatinn.

WO 90393 The finding of the synergistic response is particularly surprising given that, as shown , BHCI tends to decrease energy utilinaticn, and given that phytase has been generally used to increase growth performance via. release of eu hound phosphate, rather than through improvements in AME.

U‘I The invention is particularly important in that it enables one to minimise the input costs ated with. animal production. Specifically, as described herein, the ion. enables the minimisation of nutritive or non nutritive components without minimising the metabolisable energy of feed, and importantly, without impacting on growth performance. A tive entrapment” or “nutrient mnymnem” generally refers to an ingredient of animal feed that imbues. the feed with a particular calorific value, -nmriemr’l such as atarchee, ns, fats, oils and fibres generally provide the hull: of the energy of an animal feed, “il/Iicrrmnrrientr” generally have minimal inherent calorific value, and their core function is to enable the metabolinm of the mecronutrients. Examples of micronutrients include ns: minerals and amino acids Thus in one embodiment there is provided an animal feed including: - a nutrient component including one or more of a carbohydrate, fat and protein; —. a phytase; and - BHCl The feed is characterised in that the EEC]. and phytasse are? provided in the feed. in a ratio of amounts of BHCl : phytasc no that the AME value of the animal feed equals the AME value of I‘d (:3 the nutrient component in the presence of the amounts. of BHCl and phytnse.

According to the invention it is the synergy arising from the ratio of amounts of BHCI : e in the presence ot‘the nutrient ent, from which the AME value of the feed arisen.

More specifically, while the BHCI and phytase do not of themselves have an inherent calorific value relevant to the animal feed, it is the presence of Bl-lCl and phytase in the animal feed which increases the metabolisahle energy of the nutrient components of the animal feed.

Specifically, in one e disclosed herein“. a control diet supported an AME of 1233 Mil/kg.

Individually, BHCl addition increased this by 0.44 M] to 12.77 Nil/kg, phytase addition .margineily decreaeed this by 0.02 M! to 12,3]. Ml/‘kg, While in combination“ .BHCI addition with *pliytase addition ted an increase of HO Mlx’kg (13.43 voraus 2.33 M’Jfkg), which is indicative of a synergistic renpons‘e in energy utilisation.

The BH‘Cl and phytase are provided. in the animal feed in istioally ive amounts. ‘Synergismolly (floating amounts" of these components are amounts that. provide for an energy utilisation (or AME) that is greater than the sum of the energy ation in a feed that includes phytase only, or BHCI only. Therefore, in one embodiment there is provided an animal feed including: - a nutrient component including one or more of a carbohydrate, fat and protein; — a phytase; and l. f) - BHCI, wherein the phytase and, BHCI are provided, in the feed in synergistically effective amountg.

In one enibodin'ienn the BHCI and phytase are provided in a ratio of about: 0.53 to 5g BHCl/ltg ot‘fized : 100 to 5000 FTUfkrg offend.

‘FTU’ refers to ‘phytase unit”. Ono FTU istho ty of phytaso mquirod to liborato 1 nmol of nic phosphorus pot minute at pH 5.5 from an oxccss 0F .15 M Sodium phytato at 37°C, ably the .BHCl and phytane are provide in a. ratio of about: lg to 3g BHClll-(g of feed : 300 to 2000 FTU/kg of feed. hfl‘ore preferably, the BHC] is provided in an amount of more than 20 to about 2V7ngk‘g of feed, more pretérably. about 205g, or 2.1g or 3.253,, or 2.3g, or 2.4g or 2.5g, or 2.6g, or 2.7g/kg of food, The phytase may be provided in an amount of more than lOOOFTU to about 3000FTU/kg feed, for example from: about l,lOO FTU or, 1,200 FTU on, LBOC‘: FTU or, 1,400 FTU or, LSQO FTU or: 1,500 ,FTU or, 1,700 FTU or, 1,800 FTU or, 15300 FTU on 2,000 FTU or. 2,100 FTU mu: or, 2,200 'FTU or, 2,300 FTU on, 2,400 FTU or, 2,500 FTUi’kg feed.

In another embodiment, the phytase may be provided in. an amount if from 500 F'lU, or 600 FTUJ, er 700 FTU, or 800 FTUE, nr 900 FT’U, or 1,000 FTLl/kg of feed. In this emhedimentg the LBHC’L may he provided in an amount of from 2.05331 er 21g er 23g, or 23g, er 2.4g or 2.5g, or 2659;, or 2.7gfkg of feed.

In a particularly preferred embediment, the phytase is provided in an amount of about 50017Til/kg ef feed and the BHCI is provided in an amount at about ,2” g feed.

An amount of 300 to 2000 FTU/kg 0E feed would generally equate to about l00 tn 500g ‘phytase’tenne of feed, altheugh of course this is dependent on the concentration enzyme.

As bed herein, the invention enables the minimisatien of nutritive components 0f l0 animal feeds without impacting on energy utilisation 0r gmwth performance. For example, the invention s the formation ef feeds that have amounts of ydrate, fat or n cempenent that are less than the amount of these components that are conventionally used in specific animal feeds. By way of example, poultry feed normally contains starch in an amnunt of 300 to 5009115.; feed, pretein in an amount of .150-250gjkg, fats or oils in an amount of 50 tn SOg/kg and 100—] SOg/kg fibre. Other mieronutriente include NaCl, lysine, methionine, threonine V, limestone and inorganic phosphate, enzymes, and n and mineral premix.

According to the invention, the amount «of components of feed may he reduced so that the AME- of the nutrient eenrpontent 0f the feed (ie in the e of the hytase combination) is r than QMJIkg of feed and no more than about .1 lMJlkg ef feed, preferably eheut l0 hflfkg ef feed to less. than about llML’l-zg of feed. This represents an AME reduetlon 0f aheut l tn 2 Mlli’kg of the nutrient cempenentts‘, otherwise expressed as about a 4 to l0% reduction.

In one embodiment, the nutrient component (if the animal feed of the ion includes: - starch in an amount nf not mere than abnut 270g/kg feed? fer example in a range of from 15 t0 270g/lrg feed, — protein in an amount of lSGaZSngkg feed —‘ fats or oils in an amount of 50 to SOgKkg feed and — fibre in an amount (if 100— l50gfkg feed.

[In a. particularly preferred embodiment there is provided an. animal feed including; WO 90393 — a nutrient wmpnncnt including one at more cfa carbuhydratc, fat and protein; — a phytase; and - BHC], wherein the phytase and BHC] are provided in the feed in syncrgistically et‘fnctive U1 amounts, and wherein the nt component includes starch in an amount cf not more than abnut ZTngkg feat for example in a range of from 1 S to gc feed, In another embodiment? the nutrient compcncnt of the animal feed at the invention includes: - starch in an amount of 300 to SOOgI'kg feed - prctcin in an amount 0f I SO-ESDg/kg feed - fats or oils in an amount nfnct more than about 45gchzg feed and ~ fibre in an amount at 1 00-1 Sflgikg feed In a particularly preferred embodiment there‘ is provided an animal feed including: - a nutrient component including one or more of a carbohydrate, fat and protein; - a phytase; and - BECL, wherein the phytase and BBC] are provided in the feed in istically effective amcunts, and wherein the nutrient ent includes fats 0r oils in an amcunt ct‘nct more than about 45gx’kg feed, preferably about 20g to 45g/kg feed.

In one cmbodimcnt, the nutrient component. of the animal feed ot‘thc inventinn includes: - starch in an amount cf 300 to SOOgIkg feed — protein in an amount of ISO—ZSOgIk'g feed — fats or oils in an amount of 50 to Sfigfkg feed and - fibre in an amount of IDO—l SOg/kg feed, the component characterised in that it. does not contain one or more of the following A... :3 mieronutrients as an additive or synthetic component: methionine, choline, lysine, threonine, and inorganic phosphate.

In a particularly preferred embodiment there is prov‘i (led an animal feed including: - a nutrient component including one or more of a carbohydrate, fat and protein; _. a phytase; and - ‘B H‘Cl, wherein the e and BHCl are ed in the feed in synergistically effective amounts, and wherein the nutrient component does not include utrients as an additive or Synthetic component such as» methionine choline, lysine, iner and inorganic phosphate. [5 Further to the above, the invention enables the minimisation ofnon nutritive components of animal feeds, again t impacting on energy utilisation or growth perihrmanee, The decrease in the amount of phytase required according to the. invention compared with conventional phytase usage is significant from an economic. perspective For example, a reduction in the amount of phytase equates to a reduction in cost of'up to about 33 ftonne offeed.

In those animal feeds where phytase is not need, it is necessary to supplement the feed with phosphate because much of the phytnse derived from carbohydrate cannot be released by the animal. The invention is; larly ageous in this context because it minimised the cost input of both the amount of e required an enables one to minimiae or avoid providing free phosphate.

The invention is applicable to the prnduction of a variety 0f animal feeds, and especial!y to feeds for mnnogastrie animals and livestock rather than to ruminant animals. Examples include poultry: pork and aquaculture, Poultry feed is a particularly preferred example of animal feed. As described , U‘I poultry diets are composed primarily {if a mixture of several i‘eedstuffs (remaining nutritive components, examples of which include cereal grains, soybean meal, animal by wproducts (such. an blood and bone), fats and mineral and vitamin premixes. including anhydrnns betnine, In one embodiment the nutrient component, phytase and BHCl are combined tn form a ition suitable for ption by an . Examples of suitable compositions include granules, pellets and the like In another embediment the nutrient ent and phytase are combined to form a enmpogition suitable for consumption by an . The iti on may not include BHC]. in this embodiment, the BHCl may be prrwided tn the animal as a te composition Le. separate to the composition including. the nutrient component and phytase In nne example? the BHCl may be prnvided in drinking water.

In another embodiment there is provided an animal feed premix said premix consisting - a nan—nutrient component; - wherein the nonmutrient component includes BHCl and phytase, ‘30 Preferably the BHCl and phytase are provided in synergistically effective amnnnts.

Mere preferably the non-nutrient component further es one or mere components selected from the group consisting of an enzyme, a pigment, a growth factor, an anti —mierobial agent, such as an antibacterial. compound for inducing or ing growth performance: and an anti-mecidinl agent.

In another ment there is ed an animal feed premix, said premix consisting ~ a non~nutrient nnmpnnent; — a micronutrient component; - wherein the non-nutrient compenent includes BHCl and phytase.

More preferably the non-nutrient component r includes one or more oemponents ed from the group consisting of an enzyme, a pigment, a growth factor! an anti —nticrobial U! agent, such as an antibacterial cempound for inducing or enhancing growth mance.

Preferably the nticmnutrient component includes a ent selected from the group consisting of a Vitamin, a mineral and an amino acid.

The animal feed premix according to the invention may be provided in the form of a solid or liquid. Preferably the premix is provided in the form of a, solid, such a3 a. e or a pellet. '10 In a preferred embodiment of the invention, the phytase is of bacterial origin preferably having an amino acid sequence ofan Eeoli phytase.

Example Objective The present study was designed to evaluate the inclusions of ous phytase and betaine hydrochloride, individually and in combination, in broiler diets with three tiers- 0f nutrient specifications.

The composition and nutrient specifications of the three basal starter (1-16 days post- hatoh) and finisher (17—37 days post—hatch) diets are shown in Table it An NSF-degrading enzyme (Econese XT) was included across all the wheat-based diets to net] eat standard practice and the: r diets contained 2.0 g/kg Celite as an acid insoluble ash dietary marker The starter diets were fed as. mashn while the finisher diets were pelleted at a eonditiening ature of 85°C. The NCl diets were the ‘phytase—modified’ diets with formulated with reductions of l4 gz’kg Ca. and .l.5 gfkg P in the starter diets? which mainly stemmed from the elimination 0f diealeiunt ate, The corresponding reductione in the finisher diets were 1.2 g/kg Ca and l.4fk.g P, The NC: diets were the chetai‘ne-modified” diets in which energy density: methionine and choline levels were reduced mainly by lower inclusion levels of canola oil. and synthetic methionine and. the elimination of e chloride relative to the PC- diets. The energy densities were reduced. from 12.55 to 12.24 lekg in the starter diets and from 12.97 to 1.2.66 ,19a4 MJ/‘kg in the r diets. Methionine levels were reduced from 4.44 to ‘3, l9 talks: in the starter diets and from 4,18 to 2.93 gdtg in the finisher diets. Exogenous phytase (Quantum® Blue; AB Vista.) was included in the nt. diets at 100 g per tonne or 500 F'I’Uikg phytase activity and: similarly, hetaine HCI {Hi BetaTM s70 gl’kg betaine hydrochloride; Rural Chemical Industries) was included in the diets at 2.75 gi’kg.

The e-supplemented, experimental diets were analysed for betaine HCI contents by ometry (Appendix I). The data indicates that the six heiaine-supplemented starter diets contained an average of 3.727 g/kg hetaine HCl the six, hetaine—supplemented finisher diets contained an average 013.943 giltg hetaine HCl. The ‘baekgmund’ dietary hetaine ll} levels were not determined; however, it appears that dietary levels of hetaine HCl were higher than intended. All experimental diets were analysed for phytase activity by AB Vista using a modified ELISA method (Appendix H). The data indicates that the six phytase-supplemented starter diets contained an average of (:39l FTUng phytase activity and the six phytazse supplemented finisher diets contained an average of 589 FTU/kg phytase activity. The non- ‘Jl ‘phytase supplemented diets all contained less than SD'FTLh’kg phytase activity.

The parameters evaluated included growth performance (weight gains, feed intakes, feed sion ratios; FCR) from 1 to 163 17 to 37 and l to 3? days post-hatch. 1 to 37 days weight gain-corrected FCR, mortality/cull rates, percentage toe ash, nitrogen (N) excretion and nutrient utilisation. Nutrient ation included nt metabolisable energy (AME), expressed as both. MJi’lcg and Mil/day, N ion and N-correoted AME . Also, breast weights in absolute and relative (5% yield) terms. abdominal fat pad weights and pH of gizzard ts were determined. However, for reasuns of practical expediency? absolute breast weights were determined on a “hone—in” basis from birds that had not been tie-feathered and this approaeh inflates both the absolute weight and yield; nevertheless, the data remains indicative. The b3 experimental data was obtained and ated via standard procedures ed by the Poultry Research Foundation. While the various procedures are not detailed in this report the majority of them have been described in detail (Selle et al, 2003b) previously.

The experimental data was statistically analysed as a 3X2K2 factorial array of dietary treatments. That is diet type (PC, NC. l without and with the addition of phytase= , NCE), DJ without and with the addition of betaine .HCl. Each of the 1?. dietary treatments was offered to 8 replicate cages of 6 birds or a total of‘96 cages and S76 birds (male Ross 30?? ), The IBM® S'PSS® Statistics 20 program was used to analyse experimental data and the study eon] plied with c ines of the Animal Ethics Committee of Sydney University.

Results The effects Of dietary treatments on growth performance frern l to re days pest-hatch are strewn in 'i‘able 2 where there were no i cant interactiens between main effects. The weight gain, feed intake and feed efficiency of birds Offered PC diets were superior (P <1 0.005) to those on NCl and NC2 diets. For example the weight gain on PC diets (444 g/hird) was E; higher than NC! diets (403 g/bird) and 16.8% higher than NC2 diets (380 gibird). Betaine HCI significantly enhanced feed efficiency by 3.77% (1.380 versus 1.434; P «i 0015) in the starter phase and phytase tended to improve weight gain by 4.68% (416 vergus 4.0l g/bird; P <‘ 006) that closely approached icance. e, individually or in tandem with betaine HCI, 1 0 improved, average weight gain, feed intake and feed efficiency by 13.1,? 7.44 and 5.26%, respectively? relative to the NCI control diet. and the e FCR (1.387) was very comparable t0 the PC central diet (1.381}.

The effects (if dietary treatments on grewth performance from 1‘? to '3'? days post—hatch are shown in Table 3 where there were significant two-way interactions between all main effects ‘Jl for weight gain but not for the other parameters; The addition of e t0 the NC] diet signifi handy increased weight gain by 10.8% {1827 versus lti49 gr’bird) but increases in the PC (4.18%) and NCZ ('1..16%) diets were of more modest magnitudes. The addition (if betaine HCi to the NC} diet did not. influence weight gain (1737 versus 1739 g/hird) but. hetaine significantly depressed weight gain when added to the PC and NCZZ diets by 6.35 and 6.27%, reapectively. 2.0 dually, betaine HCI significantly depressed weight gain of the pplemented diets by 7.70% (13’97 versus 1947 } but in tandem with phytase there was no difference in weight gain (2.953 versus 1947’ gfbird) thus the combinatinn of phytase and betnine HCI ed in a 8.68% weight gain improvement in cempari son to betaine HCl en its own (1953 versus 1797 grhird). Taking, the main effects in i sedation, the. weight gain of the NCZ diet wag significantly b3 inferior to the PC and NCZ diets by 14.3 and 12.8%3 respectively. .Phytase significantly increased weight gain by 5.07% but betaine depressed weight gain by 4.489%. The feed intake of NC} diets was significantly inferior to both the PC and NCZ diets by 7.60 and 7,9l‘i-“n, respectively, and phytase significantly increased feed intake- by 335% (3286 versus 3161 g/bird; P < 0.02). The feed cenvereien ratio of NC] diets was significantly inferior to both the PC and NCZ diets by DJ 7.91 and 5.52%? respectively; and betaine HCI significantly depressed feed efficiency by 2.469%: (L707 versus 1,666; P <2 0.02).

The s of dietary treatments en growth performance from 1 to 37 days post are shown in Table 4 and, again, there were significant interactiOns n main effects fer weight gain but. not for feed intake and feed ccnversicn ratios. il’hytase tiacticnally increased weight gain by 0.97% in NCZ diets and by 3.71% in PC diets, which was significant. Hewever, phytase addition tn N01 diets ed in a significant increase with a greater magnitude of 10.6% (2249 versus 2. 33 gibird). Betaine HCI significantly depressed weight gains of both the PCS and NCE diets by 4.90 and 5.29%3 respectively. In contrast, however, betaine addition tn MCI diets fracticnally improved weight gain by 0.61% (2437 versus 2147 g/bird). Interestingly. the combined inclusion of betaine IHCl and pliy‘lase, in comparison to betaine HCi alone, resulted in a significant improvement in weight gain of 7,7193% (23 74 versus 2202 gfbird).

The effects of dietary ents on gain—ccrrected feed sinn ratios, mortality/cull rates in 37 days pest-hatch. bone mineralisation {1% tee ash) and, N excretien are shewn in Table . There were no significant interactions between main effects for gain-corrected FCR; however. each cf the main effects did have significant impacts. The gain—cerrected FCR for PC diets of 1.521 was significantly better than NCZ diets (1.616)by 5.38% and NC] diets {1.769) by ltli..3‘i»b.

Also, NC2 diets were significantly better by 8.65% than NC]. diets. Pliytase significantly enhanced gain-corrected FCR by 3.712% (1.604 versus 1.666; P <1 0.03); however, betaine HCl significantly depressed gain-corrected FCR by 3.42% {1.663 versus 1.608; P < 0.05). The overall ntcrtalityfcull rate was 4.51% but was unrelated to treatments. Phytase increased toe ash by .95% (1.2.4? versus 1.1..76‘3-ti; P <1 0.01), which was the enly significant. effect. ed in. respect of bone mineralisation. Betaine HCl d N excretion by 7.6996 (38.79 versus 42.02 2.0 d; P <1 0.005), which was the only significant effect observed. However, phytase numerically reduced N excretion by 3. Bit-i) (39.59 versus 41.2.] g’bird; P < 0.15). Interestingly, across all three diet types, the lowest N excretion. rates were nbserved in diets supplemented with bath betaine HCl and e.

The effects of dietary treatments on nutrient utilisatinn are shown in Table 6. Significant {*4 U1 interactions were observed for AME (Ml/’kg) between diet type and b‘etaine additinn (P '< 0.05) and between additions of betaine and phytase (P <1 0101). The on of betaine tn NCI diets increase-d AME by 0.78 MJ (1310 versus 12.32 ); Whereas; additions tn PC diets resulted in a ccmparatively modest increase of 0.20 M] and to NC2 diets a slight decrease of 0.08 M]. in the absence of e, betaine addition resulted in a numerical se of 0.08 M]. —writ= DJ (3 in the ce of phytase. betaine addition generated an se of 0.67 MI (1368 versus 1.3.01 lit-11.11113). Taken separately, there were significant outcomes for the three main effects of diet type (P < 0.001) and alsc phytase and betaine additions. NC} diets (12.71 M‘Ji’kg) had significantly lower energy densities than either PC (13.53 g) cr NCZ (13.30 Mill-{g} diets. Pbytase increased AME by 0.34 Mllkg (1335 versus 1301 Mllkg; P < 0.02:) and betaine increased AME by 0.30 Mllkg (13.33 versus 13.03 Evil/kg; 'P <1 0.04).

One signifi cant. interaction was observed for AME. (“.Ml'fday), Which was between additions of betaine and e (P “I. 0.005).. In the absence of phytttse= hetaine addition depressed energy intake by 0.115 Ml or 5.43% (2.002 versus 2117 ). However, in the presence of phytase, betaine addition enhanced energy intake 0.120 MI or 5.63% (2.251 versus 2.13.1 MJz’day). Taken separately; both diet type and phytase addition significantly influenced energy intake; whereas, betaine did net influence this parameter (P 43* 0.90). The energy intake of Mill diets (1.940 y) was inferior to both. PC 2.234 .Ml/day) and 'NCZ (2.200 Mllday) diets. 'Phytase additien increased energy intake by 0.132 Ml/day or 6.41% (2.19:, versus 2.059 .MJ/dny; P < 0.001).

Overall, the birds retained 52.12916 N; however, there were. .110 significant treatment effects observed. for N retention. There were two significant interactions between main effects for N-corrected AME; these were for diet type K hetaine HCl (P <1 0.01) and phytase x betaine HCl (P < 0.03). The on of ‘betaine HCl to NC] diets increased AMEn by 1.01 Ml {11.36 versus 1067 hill/kg), which was considerably more pronounced than the corresponding ses in PC (0.23 MJ} and N62 (0.17 Ml) diets. In. the absence of phytaset. e .HCI sed r—XLMEn by 0.20 M] (11.46 versus 11.26 Mlx'lrg). liloweyer, in the presence of phytase, betaine HG] increased AMEn by 074 Ml (12.13 versus 91V1Jfkg), which is also a erably more 2.0 pronounced se. Taken in isnlatinn, all three main effects had significant impacts on AMEn. The Mill diets were cantly inferior tr) both PC and NCZ diets. Phytase increased AMEn by 0.40 Ml or 3.52% (11.76 versus 1.1.36 Mllkg; P < 0,005) and e H'Cl inc-teased AM‘En by 0.46 M1 or 4.06% (11.79 versus '1 1.33 lekg; P ’4. 0.001) The effects of dietary treatments on absolute and relative breast weights, fat pad weights and gizzard pH are shown in Table 7. There was a significant interaction (P < 0.005} between diet type and phytase addition for absolute breast s. Phytase increased breast weights by "12.9% in NC: '1. diets (603 versus 534 g/hird) but the imprtwement was a. mere modest. se nt‘ .837% in PC diets (703 versus 664 g/hird) with no real difference in NC? diets (628 versus (334 gfbirdf). As main eft‘t—ictsa there were significant differences between PC (683 ), NC? (630 DJ (3 gfhird) and NCl (5:63 gfhird} diets. Also phytese significantly increased absnlute breast weights by 5.57% (644 versus 610 g/bird; P < 0.001) but hetaine addition had no influence (P > 0.40).

There was a significant interaction (1’ < 0.015) between phytase and hetaine additinns for relative breast weights or percentage yield. In the absence (if phytase, betaine HCl increased. yield from .9 to 27.0%, but. in the presence of phytase, hetaine HCI fractionally decreased breast yield from 26.5 to 26.49523. The main effect of diet type was significant (P {I 0.001,) where the yield on PC diets (27.4%) wan superior tn NC] (25.9%) and NCE ) diets, Betaine HCi increased breast yield by 0.5 percentage units (26.7 versus 26.2%; P < 0.05) but. phytase had no influence (P T» (3.90) on this parameter.

There was a significant interaction (P < 0.005) between phytase and betaine additions fat fat pad weights. In the absence of phytase, betaine HCl decreased fat pad weights by 17.6% (16.8 versus 20.4 gz’bird), but, in the presence ot‘phytase, betaine 'HCl increased fat. pad weights by 8.67% {2.1.3 versus 19.6 g/hird). The main effect. of diet type was significant {P < 13.001) ll} where fat pad weights on NCI (116.6 gfbird) diets were less than both PC (20.3 gfbird‘) and NCZ (2.1.7 gfbird) diets. Phytase sed fat pad weights by 10.294; (2.0.5 versus 18.6 ; P < 0.04) but hetaine HCl had no effect (P 3} 0.25:) on this parameter.

The pH of digesta in the gizzard was determined for birds offered PC and NCi diets and was analysed as a 2x2x2 factorial array of treatments where there were no cant ‘Jl interactions between main effects. Gizzard pH of NC]. diets was Significantly lower than PC diets (3.46 versus 3.6]; P < 0.65); however, pbytase had n0 effect (P > 0.50) on gizzard pH. In eentrast, e HCl significantly reduced giezard pH by 0.28 (3.39 vetting 3.67; P < 0.005). sian The 2012 Aviagen performance objectives for male Rose 308 chic-its at 37 days post- hatch is for a. body weight of 2457’ g, a feed intake of 3944 g and an FCR of" 1.605. If the 42 g day—old chick weight is deducted this translates to a weight gain 5 g and an FCR of 1.633.

In the present study, birds offered the nan-supplemented. PC diets had. a weight gain of 2530 g and an FCR of 1.591, which represents respective improvements of 4.76% and 2.57% ve t0 the perfnmiance objectives. ii) As tabulated, meat~and~hone meal was anaiysed to contain 4.1.7 g/ltg P and it is d that dicalcium phosphate ned 180 gx'kg P. Samples of wheat and soybean meal were analysed by an al laboratory and were ed to contain 250 and 6,65 gz’kg total P, respectively. U‘nt‘ertunately, however, the phytate—P levels reported were not credible and these values were estimated train Ideally generated data. (Selle et 31., 2003b) en the basis et‘tetal ’P DJ enntents because the twe parameters are significantly correlated. On this basis, wheat and soybean meal contained estimated levels. of 1.85 and 4.53 g/fkg phytatei", respectively. Thus the estimated total P, phytate—P and nenphytate—P trntinns nt‘ the Six basal diets are Shawn in Table i; on average, the diets centained 4.34 gi’kg total P, 2.45 gfkg phytate-P and 1.85 gfltg nonphytate—P. Thus, there was a moderate substrate level of 8.69 gx’kg phytate or 2.45 giltg phytate—P. However, the estimated. e levels of434 giltg total P and 1.85 gfls‘g nonphytate- P, especially the latter, do appear suspiciously low given the growth performance of the birds d these diets. The reported 2.50 g/kg total P concentration in wheat does appear low as Selle et al. (2003b) reported an average total 'P t 8 ge’kg in 37 wheat s.

The estimated nonphytate-P levels in the NCI starter and finisher diets of 1.15? and 1.825 going, respectively, are noticeably less that the specified available P levels of 2,50 and 2.00 git/”kg respectively, Therefore, it is noteworthy that. toe ash was numerically lowest in NC 1 diets and, overall, phytase Significantly increased toe 3511 by 6.64% (12.47 versus 1 1.76 gr’kg; P <3 ll} 0.01), Moreover, specifically in the N01 diets, phytase Supplementation increased average toe ash by 8.66% (12.42 versus 11.43%). The likelihood is that. the diets were limiting in either nonphytate‘ or available 13., which would amplify responses to phytase; whereas, as would he expected. betaine HG]. did not have a significant influence on toe ash 7 versus 12.15%; P 3> 0.75).

‘Jl In the discussion that follows the initial focus is on the impact of diet type; mainly on growth performance. Secondly, the main effects of betaine HCl and phytase on the ters assessed will be considered, followed by assessments of the interactions between the dietary inclusions of phytase and betaine HCI.

From 1. to 37 days post-hatch, weight gains of birds offered the non-supplemented NCl diet was 13.9% less than the control PC diet {2053 versus 2530 gihird) but phytase plus hetaine ‘HCl Supplementation of'NCl diets ted a l 1.2% increase in weight gain (2282 versus 205.3 gfbird) so that the deficit to the PC diet was 9.8%. Feed intakes followed a similar pattern with a 13.2% reduction with the transition from PC to NCl control diets (3495 versus 40:25 d) and phytase supplementation of the NC 1 diet ted a 7.9‘2’6 increase in feed intake (3?70 versus b3 349.5 ghird). The FCR of l. PC diets was 1.591 and the FCR of control NC.1 diets was 1.104, which represents a 710% deterioration in feed efficiency; however phytase plus betaine HCl supplementation of the NC] diet. increased feed efficiency by 3.81% (1.639 versus 1.704,).

Thus phytase supplementation of NC] diets partially compensated the reductions in growth performance parameters generated by the transition from PC to Mill control diets. That this DJ sation was partial may be attributed to very low non-phytete P levels in the NCI diets coupled with relatively modest levels of the substrate, e.

The reductions in performance parameters arising from the transition from PC to NCZ l diets were considerably more modest with reductions of 3.2%, 2.8% and 0.4% in weight gain, feed intake and FCR, respectively. The inclusion ot‘hetaine HICI and *phytase in. NC 2. diets ted a smell 1.1% increase in feed intake (3955 versus; 391 1 g/bird) hut hetaine HCl and phytase, singly and in combination, did not e either weight gain or feed conversion ration supported by NC2 diets.

The influences of diet type and supplementation. on N«corrected AME are of interest. The transitinn from l PC to NC‘l diets reduced AME-n by 9.4% or 1.1! MJng (1070 versus 1.1.81 MJ/kg). However, phytase plus hetaine HCI supplementation of NC ] diets increased AMEn by 13.1% or 1.40 MI or ("12.10 versus 10.70 hill/kg) resulting in a higher AMEn value by 0.29 Ml (2.46%) than the control PC diet, which is a noteworthy outcome The transition from 1 0 PC to NCZ control diets reduced AMEn by 4.49% or 0.53 MJ (11.28 versus 11.8] Mii’kg'); however, e plus betaine HCl supplementation of NC‘Z diets increased AME by 6.29% at 0.71 Mil/“kg (l 1.99- versus 'll.281\fIJ/itgj, which was 0.18 MJ (1.52%) higher' than the control PC: diet. Thus e and hetaine HCl in tandem highly were effective in enhancing nutrient utilisation as esgesscd by Ext-corrected AME.

‘Jl As growth mance main effects, phytase significantly (P <1 ) increased weight gain by 5.07% in the finisher phase and by 4.84% from i to 37 days post-hatch. in addition, e significantly (P < 0.01) increased feed intake by 3.76% over the entire feeding period and also significantly (P < 0.03) improved weight gain-corrected FCR, by 332%. This finding is noteworthy as positive growth. performance responses to se are often confined to 2.0 weigh gain and feed intake rather than efficiency of feed conversion. Phytnse supplementation of NC 1 diets generated robust improvements of 10.6% in weight gain, 7.63% in feed intake and 2.79‘3’6 in feed conversion from 1 to 37 dayg poet—hatch.

As nutrient utilisation main effects, e significantly increased AME by 0.34 lelrg (P <1 0.033) and 0.132 MJ/day (P < 0.001) and Al‘v‘fEn by 0.40 Mir'kg (P < 0.005), which are b3 tangible ses in energy ation. Somewhat curiously. the 3x2x2 factorial array of treatments, including phytase, did not significantly influence N retention. Phytase significantly increased (P <1 0.04) abdominal fat pad weights by 10.2%, which may be an indicatnr of enhanced energy utilisation.

The significant main effects of betaine HCl on growth performance included an DJ improvement of 3.777% in LFCR in the starter phase huti alternatively. a depression of 2.46% in the fininher phase such that there was no significant impact (P >- 0.30) from 1, to 37 days post— hatch, However, betaine Hill Significantly depressed growth rates in the finisher phase by 4.48% and by 3.38% over the entire feeding period, which suggests the 2.75 gr'kg inclusion rate may have been ive.

Of interest is that e HCl significantly (:1) 4' 0.005) reduced N excretion by 7.69% from 42.02 to 38.79 gfbird during the total excreta collection period. This is an important outcome given the s environmental implications. Also. as a main effect, betaine HCI significantly ed energy utilisation by either 0.30 MJfkg as AME or 0.46 MJz’kg as AMEn.

Importantly, hetaine HCl significantly (P < 0.05) increased breast meat yield by 1.91% from 2.5.2. to 26.7% as a main et’f’eet. Finally, betaine Hill significantly reduced (P < 0.005) the pH. of gizzard contents from pH 3.6?“ to 3.39, which may age the ty of the E. Coil—derived phytase given its pH activity spectrum.

The significant interactions observed between the dietary additions of e HC] and e are of particular relevance. There was a significant interaction (P = 0.003) between betaine HCl and phytase for weight gain from i? to 37 days post-hatoh. Betaine HCl alone (2.943 gi’kg analysed} depressed weight gain by 7.70% (1797 versus 1947 g/bird) but in the ‘Jl presence of phytase (58??? .FTUYkg analysed) there was a fractional increase in weight gain of 0.31% ([953 versus l947 g’kg). Thus the use of phytase and hetaine HQ] in tandem sed weight gain by 8.68% 0953 versus 13’97 g/bird) relative to the use of betaine HCi alone. In on? there was a significant interaction (P = 0.009) between hetaine HUI and phytase for weight gain, with a similar outcome, from i to 37 days post-hatch. The inclusion of heteine HCl 2.0 at 2.75 gricg notionally may have been excessive and, if so, the analysed concentrations (3.727 and 2.943 ) wetild have exacerbated the situation. Nevertheless. it appears that phytase attenuated the negative impact of possibly excessive betaine HCl inclusion levels.

Significant interactions involving nutrient utilisation were also observed between hetaine HCl and phytase for AME BpreSSEd as MJi’kg (P 3 0.009) and MJ/day (P = 0.0M) and N- b3 eorrented AME (P : 0.022). For example, betaine HCl alone slightly depressed AME (hillfkg) by 0.08 Ml but in tandem with phytase there was a substantial increase in AME of 0.67 leltg.

Thus the betaine HCl and phytnse interaction for AME followed the same pattern that was in evidence for weight gain. in respect of carcass traits there were significant. interactions between hetaine HC1 and DJ phytase for breast meat yield. (P = 0.011) and. abdominal fat pad weights (P = . Here e HCI alone increased breast meet yield from 25.9 to 27.0% but in combination with phytase there was a frantic-rial depression in yield from 26.5 to 26.4%. Also hetaine HCII alone decreased fat pad weight from 2.0.4 to 16.8 gfbird but in oombination with phytase there was an increase from 19.6 to 21.3 gl’bird, ’li‘nus the interactive patterns for theee two carcass traits were quite different for tho-5e nhserved between betaine HCl and phytase for weight gain and energy utili satien.

It is instmctive to compare the tandem inclusion of phytase and hetaine H.121 with their individual inclusions. For example, far l to ‘37 days post—hatch weight gain the tandem ion ted a weight gain of 2282‘ g/bird in NC] diets. This; was 3.02% higher than phytase alone (221.5 gfbird) and 3.42% higher than e HCl alone (2012 g/bird). Similarly, phytase plus hetajne HCl supported a Feed conversion ratio nt‘ '1 {639 in. N‘Cl diets ever both feeding phages.

This repreeented improvements in. feed efficiency of 3.64% ve to phytase alone (1.701) and ll} an improvement of 5.37% relative to ’beteine HCI alone ).

Of real interest is the effect of dietary inclusions in NCZ diets- on N excretion, Birds offered NCZ control diets ed 45.68 g/bird N. Betaine HCI reduced this figure by 7.03% to 42.47 gfhird and phytase reduced this figure by 10.7% to 40.80 g/bird. However, the combination ef phytase and betaine HCI generated a fully additive reduction in N excretion of 18.9% (37.05 versus 45.6% gfbird). y, the effect of dietary inclusions on AME (MJKkg) in NCl diets is considered. The central diet supported an energy density 0? 12.33 Mllr'kg. Individually betaine .HCl increased this by 0.44 M] to 12.77 MJ/Tltg and, somewhat Sumrisingly, phytase marginally sed this by 0.0: MI to 12.31. . However, in. combination? betaine HCl plus phytase generated an increase of 1.10 MJ/kg (1343 versus. 1233 MJfkg), "which does appear to be a synergistic response.

References: Selle PH at at, 2003(311An51. J. Expert Agricul. 45 :475-479 a"21 Table l Compasition and nutrient specifications of basal diet-s ge (0.100 g/k’g) and e HCI (2.750 ) were added to the appropriate diets at the expense (if wheat] Starter dials Finisher diets NC 1 Mill NC} Wheati 638.3 639. 10 1348.5 Soybean meal ($18)" 310.0 299.0 253.4 Meat and bone meal (50)} 22.0 30.0 30.0 Canola oil 17.0 6.0 213.0 Limsstone 9.0 6.0 6.0 Dicalcium phosphate 0.0 8.0 4.0 Sodium chloride 3.2 3 .6 2.0 Sodium bicarboimte 2.0 2.0 2.0 Lysine HE] 2.4 2.5 2.4 Methionina 1'" [‘0 0.9 2.2 Threonine 5' . '4’) 0.3 0.5 Choline chlaridc (60} —‘ 3 01) 0,4 0.4 Viimnin-mincral premix 1'9 U1 2.5 2,3 2.5 Ewnasc \T Av .— 0.1 0,1 0.1 Celine 20,0 20.0 Metabolisablc energy g) 12.5.5 12.55 12 24 1 2. _ , 9 7 12.97 Calcium 8.40 7.00 8.40. 7. 40' {1.20 Available phosphmus 4.00 2.50 4.00 3 . 40' 2.00 Sodium 2.40 2.20 2.40 1. . 80 1.80 Choline 1.85 '1 .85 1. . 5.0 1.50 L}; sine 12.0 12.0 12.0 11.0 11.0 Methimfine 4.44 4.44 3.19 4.18 4.18 ivksihimline 311d cystinrc: 8.40 8.40 7.15 8.03 8.03 Tiypmphim 1.92 '1 .92 1.92 1.87 1.87 Thrconinc 7.44 7.44 7.44 7.04 7.04 A t‘gininc 12.60 1 2 .130 12.60 11 .88 11.38 {solelicine 7.80 7.80 7.80 7.37 7.37 Vanna 9.24 9.24 9.24 8. 58 8.58 Extimmm Total P 5.131 rd») :1 4.225 £413,7me 2.520 l 1 37 2.400 Nonphytate P 2.611 L.l,.... UI’WN - 1‘41“?” Cfiflflh 51. 1-1 1.825 [Contained 2.50 gx'kg P (Symbio Alliance, lob NO 196546) and estimated 1.85 gfkg phytate—P. 2Contained (3.65 gig/kg 'P (Symbio Alliancve, Job No 196546) and estimated 4,53 g/kg phytate—P. 3'(ijiirtainacl 8.15 g/kg Ca and 4.1"? g/kg P (Grain Brewers, Job No 02191) R3 {'43 Table 2 Effect of dietary treatments on growth performance from 1 to ‘16 days past-hatch Treatmentg Growth erfbrmance Diet Phytase Retains. Weight gain Feed intake Feed ' ‘ ’ (‘1‘ 11d) (Ufbird' efficiency PC 0 0 43? 603 1.381 0- 2.75 444 5511 1.334 501.1 0 441 590 1.341 500 2.75 453 5915 1.313 NC] 11 0 373 544 1.464 0 2.75 395 556 1.414 5110 11 4'18 591 1.415 501.1 2.75 426 573 1.359 NCE 1.1 0 33:1 5711 1.503 0 2.75 377 523 1.394 500 0 378 564 1.502 5110 2.75 383 557 1.452 SEM 13.448 15.103 0.0363 Main effects: Diet type PC 4446 595b 1.3451“ N121 403b 5675" 1 . 13b 111:2 3813“ 553R 65“ Phytase 1:1 41.11 554 1.415. 5110 FTng 416 579 1.399 1361211116 0 405 57 1.434a 2:75 gx’kg 413 557 1.5311" Significame (P =) Di :1 type (D) <:0.01:11 11.111312 <: 0.1101 Phytase (P) 0.054 0.115 0.460 e 113‘) 11.313 0.279 2 D x P 11.12:; 0.252 0.252 D x B 0.72? 0.438 0.745 P x B 0.935 11.552 11.492 D x P x B 0.798 0.371 (1159 [KB 1;) Table ‘3 t of dietary treatments- 011 growth perfbrmance from 17 to 3? flags post~hatch Treatments Growth. aflrfoninance Diet 13113112136 Bis-taint: Weight gain Feed intake Feed (FTUfkgz) {WI/{U} aW’bmi j11311215) efficiencv PC 0 0 2094 3422 1.636 0 2.375 1878 3.123 '1 665 500 O 2094 3 340 1.597 .00. 2.75 2044 3328 1.627 NC.1 0 U '1 68.0 2950 1.315“?r O 2.75 1618 2923' 1.311. 500 0 1 798 3 .179 '1 .769 500 2.?5 18:37 3 .157 1.104 NCZ 0 O 2067 3342 1.617 0- 2.75 18% 3205 1.691 500 O 2047 331 I 1.619 500 2.75 1960 3 398 1.746 SEM 34.944 81.503 0.0532 Main effects: Diet type PC 2027‘” 3 3031‘ 1 631*" NCI 1738a 3052” 3 N02 1993b 3314’“ '1, .668b Phytase 0 18723 31613 1.696 soomm'kg 1967‘“ 32.861“ 1.677 Bataine D .1963” 3257 1.6661: 2.?5 g/kg 1875" 3.189 1.707h Significance (P :) Diet type (D) <2 0001 < 0.00] a: {1.001 Phytase (P) <4 0001 0.010 0.471 Bataina (B) <1 0.001 0.149 0.015} D x P 0.008 (1.273 0.451 D x B 0.013 0.428 0.252 P 3:13 0.003 0.070 0.686 D. \ P x. B 0.712 0.443 0.396 2014/050052 Table 4 t of dietary treatments- 011 growth perfbrmance from 1 to 37 days post«hatch Treatments Growth. aflrfomlance Diet Phytase Bis-taint: Weight gain Feed intake Feed (FTUfkgz) {WI/{U} aW’bmi 'flbird) efficiencv PC 0 O 2530 4025 1.591 0 2.?5 2321 3714 1.601 500 0 2536 3930 1.550 500. 2.75 2497 3924 1.570 NC,1 .0 0 2053 3495 1.704 0 2.75 2012 3478 1.732 500 0 2215 3770 1.701 500 2. "75 2282. 373 5 1.639 NCZ 0 0 2450 391 1. 1.597 0- 2.75 2272 3728 1.641 500 0 24215 3875 1.5199 500 2.75 2344 3955 1699 SEM 40.529 87,166 0.0387 Main effects: Diet type PC 2471 3898’“ 1.578.“ NCI 214l 3619fl 1.694a N02 23:73 3867’” '1, .634b Phytase 0 2273 317253 1 .644 500 .FTm'kg 2383 38653“ 1.626 Bataine 0 2368 3834 1.624 2.?5 g/kg 2288 3754?: 1.647 Significance (P :) Diet type (D) <2 000‘] < 0.00] a: 0.001 Phytase (P) <10, 001 0. 007 0.429 Bataine (B) 0.001 0.122 0.307 D X P 0.004 0.204 0.324 D X“ B 0023 0.523 0.270 P KB 0009 0.0"2 0.853- D x P x. B 0.789 0.365 0.401 [‘23 ’41.

Table 5 Efi‘éc’t of dietary treatments on gain~corrected feed conversion 172111700, mortalityfcull rates], bone mineraligation (percentage toe ash) and N excretion during 1010.1 excrata cellecfion period Treatments Gain- Mmtality corrected and 0011 Diet. Phytase Betaine FCIR rams Toe 3311 N 153pe (FTU1kg) (.31kg) («r .53) (95’s) (‘14:) ian ('gx’bird) PC 0 O 1.510 4.18 11.30 39.97 0 2.75 1.604 4.18 11.99 40.34 :00 ‘0 1.467 8.35 12.29 '3 9.59 500 2.75 1.503 2.091 12.54 39.20 NC1 O O 1.214 51.25 11.70 41.52 0 2.75 1.858 4.18 11.15 37.30 500 0 1.747 418 1262 44.5.4 500‘ 2.75 1.657 6.26 12.22 36.38 NCQ O 0 1.548 2.09 12.10 45.68 0 2.75 1.6263 13.25 12.29 42.4? 500 O 1.560 4.18 12.88 40.801 500 2.75 1.692 209 12.24 37.05 SZEM 0.0474 3.266 0.4171 1.8227 Main affects: Die-t type- PC I .521“ 4.69 12.03 39.78 NCI 1769‘: 5.21 ,1 1.92 3.0.93 NCE 1.616" 3 35 12.33 41.50 Phytase 0 1.656“ 4.51 21 41.21 500 .FTUfkg 1.0041’ 4.51 12.471 39.59 Retaine- 0 mosh 4.86 12. .15 42.02“ 2.75 gk‘g 1.6633 4.17 12.07 38.319“ Significance (P :) Diet type (D) <: 0.001 0.789 0.289 0.341 Phytase (P) 0.020 0.990 0.006 0.128 86131116 (8) 0.046 0. 713 0. 755 0.003 D X P 0.073 0.903 0.567 0.052 D x B 0.096 0.644 0.264 0.061 P X 13 0.291 0.463 0.448 0.409 D X P x B 0.532 0.431 0.?11 03131 Table 6 Effect 0f dietary ents m1 apparent metabalisahie energy (AME: M’Jfkg, M'deay}, nitmgen (N)1 111 (.111 and ected AME (AMEH) Treatments AME; AMEn D1 e1 Phytase Betaine (1»thng AME N retention {MJ’fkg 11,2 .10 (FTUI’k ) (21kg) DM) (MJXday) (‘34:) DM) PC 0 0 13.63 2.330 55.22 11.31 0 2.75 13.29 2.077 52.05 11.69 500 0 13.23 2.207 50.08 1 1. .113 500 2.75 13.96 2.324 03.72 12.30 NC1 0 0 12.33 1.1111; 50.80 10.70 0 2.75 12.77 1.868 52.14 11.25 500 0 12.31 1.953 50.55 10.64 500 2.75 13.43 2.123 49.81. 12.10 NC2 0 0 13.19 2.203 53.2 11.28 0 2.75 12.30 2.1101 49.25 11.43 500- 0 13.50 2.233 53.30 11.30 500 2.75. 13.66 2.305 55.16 11.99 SEM 0.2406 0.0570 .1 .9438 0.2025 Main effects: Diet typa 1313 15.53“ 2.234“ 52.77 11.38“ NC1 12.71“ 1.940“ 50.83 11.178 1510: 13.301- 2200” 52.74 1 1.62h Phytase 0 13.01“ 2.059" 52.12 1.1.36a 500 'FT‘Ufkg 13.35h 2191b 5211 1176b 30121110 0 13.03” 2.124 52.2 11.33“ 2.75 glkg 13.33b 2.120 52.02 11.119“ Significance (P 5) Diet type (D) < 0.001 < 0.001 0.271 < 0.001 Phytase (P) 0.018 «r. 0.001. 0.990 0.001 Betaine (3‘) 0.035 0.940 0.868 < 0.001.

D x P 0.466 0.265 0.121. 0.641 D x B 0.040 0,071. 0.851. 0.007 P x 13 0.009 0.001 0.118 0.022 D x1) x B 0.683 0.303 0.208 0.301 2014/050052 Table 7 Efi‘éc’t of dietary treatments an ab salute breast weight, breast meat yield, fat pad weights and gizzard pH Treatments Breast Breast Fat pad Diet Phytase Betaine weight meat weight Gizzard 'iype (FTUI’kg) (gxkg) (gr’bird) yield (gfird) pH (9%} PC 0 0 668 26,1. 21.8 390 0 2.75 660 28.3 17.3 3.40 500 0 708 27.6 21.0 3.70 500 2.75 697 27.5 21.0 3.45 NC} 0 0 541 25.7 16.4 3.58 0 2,75 526 25.7 14.7 3.35 500 0 581 26.0 17.1 3.51 .00 2.75 624 26.2, .1 8.3 3.38 NCZ O .0 642. 25.9 23.1 - 0 2.75 625 20.0 18.4 — 500 0 643 26.0 20.8 — 500 2,75 6112 25.5 24,7 — SEM 15.2.47 05384.1 1. .442 0.1057 Main effects: Diet iype- PC 6836 27.4" 20.3b 3611* NC 1 5683' 25 . 9;: 16.6“ 3 .463 N02 63 0b 26.13 2‘: .7b _ 0 6103 20.4 18.6a 3.56 500 .FTUfkg 644b 2M 2051’ 3 .51 Betaine 0 631 2.6.23 20.0 3671‘ 2.?5 gi’kg 624 26.7b 19.1 3.393 Significance (P r") Diet type (D) <3 0.001 <0.001 <1 0.001 0.044 Phytasa (P) <i 0.001 0.930 0.030 0.540 Betaine (B) 0.436 0.041 0.257 0.00.1 D x P 0.004 0.084 0.934 0.752 D x 8 0.2.26 0.184 0.554 0,313 P X 3 0.434 0.011 0.002 0.251 DXF‘ XB 0.210 0.070 0.359 0.584 Gaming Angofiohummmw m; imam. wwfimmm manoww mowvmfiq 322 wwbohm ,mmmm mmgmom mcvmmmm mmdmwm momma... fly m flammEuE $qu mxxwmnm mémfiw Emma 3?: mafia 33m wémmfim 3&3 3&3,qu mg‘mmnm Burn :2“: fix: a“; £5 fig :5; ii E? EE 3.5 EEQEQ $5.? .mwmfimwfi at mfiiqmfi B3: Em 5in wmfifium kfiwbmfiw 2 2Q saw Stamgm awfifiWn-w gmtsmae Egmfim 335%-?“ nmfifimfimm a afifiméu w Emmaié “Ewaaiw m mm; mfimvm 3mm; 3:3— “gum “23:52: EmEEAfl L43 C3 Appendix II Sample informatiun PhytaSe activity (QB Elisa FTUlk ) 1A r 0 FTU <1 50 28 StarterO FTU <1 50 3C Starter SOOFTU 895 4D Starter 500 FTU 790 513 Starter 0 FTU 6F r 0 FTU 78' Starter SUUFTU 455 8H Starter 5.00 FTU 567 9]? r 0 FTU < 50 } Starter 0 F177;] < 50 11K Starter SOUPTU 700 12L Starter 5'00 .FTU 737 1A Finisher U FTU 28 Finisher 0 FTU 3C. Finisher 500FTU ‘l 1% 4D Finisher 500 FTU :1 0% SE Finisher 0 FTU 6F Finisher O FTU 7G Finisher SDOFTU 8H Finisher 500 FTU 91 Finisher 0 FTU 101‘ Finisher O FTU 11K Finisher SOUFTU 12L Finisher 500 FTU 1003291242

Claims (23)

1. An animal feed ing: - a nutrient component ing one or more of a carbohydrate, fat and protein; - a phytase; and 5 - betaine hydrochloride , wherein the BHCl and phytase are provided in a ratio of about 0.5 to 5g BHCl per kg of feed : 100 to 5000 e unit (FTU) per kg of feed; and n the feed is a poultry feed.

2. The feed of claim 1 wherein the starch content of the feed is not more than 270g per kg of feed. 10

3. The feed of claim 1 or claim 2 wherein the protein content of the feed is from 150 to 250 g per kg of feed.

4. The feed of any one of the preceding claims wherein the fat content of the feed is not more than 45g per kg of feed.

5. An animal feed premix, said premix consisting of: 15 - a non-nutrient component; - wherein the non-nutrient ent includes BHCl and phytase, and wherein BHCl and phytase are provided in a ratio of about 0.5 to 5g BHCl per kg of feed : 100 to 5000 FTU per kg of feed; and - wherein the animal feed is a poultry feed. 20

6. The premix of claim 5 wherein the non-nutrient component further includes one or more components selected from the group consisting of an enzyme, a pigment, a growth factor, an anti-microbial agent and an anti-coccidial agent.

7. The premix of claim 6, wherein the anti-microbial agent is an antibacterial compound for inducing or enhancing growth performance. 25

8. An animal feed premix, said premix consisting of: 1003291242 - a non-nutrient component; - a micronutrient component; - wherein the non-nutrient component includes BHCl and e, and n the BHCl and phytase are provided in a ratio of about 0.5 to 5g BHCl per kg of feed : 100 to 5 5000 FTU per kg of feed; and - wherein the animal feed is a poultry feed.

9. The animal feed premix of claim 8 wherein the non-nutrient component r es one or more ents selected from the group consisting of an enzyme, a pigment, a growth factor, an anti-microbial agent and an anti-coccidial agent.

10 10. The premix of claim 9, wherein the anti-microbial agent is an antibacterial compound for inducing or enhancing growth performance.

11. The animal feed premix of any one of claims 8-10 wherein the micronutrient component includes a component selected from the group consisting of a vitamin, a mineral and an amino acid. 15

12. An animal feed or animal feed premix of any one of the preceding claims wherein the phytase is of bacterial origin.

13. The animal feed or animal feed premix of claim 12 wherein the phytase has an amino acid sequence of an E.coli physate.

14. The animal feed or animal fee premix of any one of claims 1-13, wherein the 20 nutrient component does not include the following micronutrients as an additive or synthetic component: methionine, choline, lysine, threonine and inorganic phosphate.

15. The animal feed or animal feed premix of any one of claims 1-13, wherein the feed is in the form of granules or s.

16. The animal feed or animal feed premix of any one of claims 1-15, wherein the 25 phytase is provided in the feed in an amount of more than 1,000 FTU/kg to about 3,000 FTU/kg; or the premix is for combination into a feed providing phytase in an amount of more than 1,000 FTU/kg to about 3,000 FTU/kg. 1003291242

17. The animal feed or animal feed premix of any one of claims 1-15, wherein the phytase is provided in the feed in an amount of about 500 FTU/kg to about 1,000 FTU/kg.

18. The animal feed or animal feed premix of any one of claims 1-17, wherein the BHCl is provided in the feed in an amount of more than 2g/kg to about 2.75 g/kg. 5

19. The animal feed or animal feed premix of any one of claims 1-15, wherein the phytase is provided in the feed in an amount of about 500 FTU/kg and the BHCl is provided in the feed in an amount of about 2g/kg.

20. The animal feed of any one of claims 1 to 4 substantially as described herein with reference to the examples. 10

21. The animal feed premix of any one of claims 5 to 7 substantially as herein described with reference to the examples.

22. The animal feed premix of any one of claims 8 to 11 substantially as herein described with nce to the examples.

23. The animal feed or animal feed premix of any one of claims 12 to 19 ntially 15 as herein described with reference to the examples.