CN104684408B

CN104684408B - Feed addictive composition

Info

Publication number: CN104684408B
Application number: CN201380051433.3A
Authority: CN
Inventors: L·罗梅罗; P·普勒斯泰德; L·巴尔纳德
Original assignee: DuPont Nutrition Biosciences ApS
Current assignee: DuPont Nutrition Biosciences ApS
Priority date: 2012-08-03
Filing date: 2013-08-02
Publication date: 2018-08-31
Anticipated expiration: 2033-08-02

Abstract

Performance for improving individual,Or digestibility (such as the Nutrient digestibility for improving the raw material in feed,Such as amino acid digestibility),Or retain for improving nitrogen,Or for improving the absorption of diet phosphorus and retaining,Or the effect of for improving phytase,Or the resistance for improving individual to necrotic enteritis,Or for improving feed conversion rate (FCR),Or for improving the weightening in individual,Or for improving the feed efficiency in individual,Or the immune response for adjusting (such as improvement) individual,Or the population of the pathogenic bacteria in the gastrointestinal tract for reducing individual,Or the method for reducing the excretion of the nutrients in muck,This method includes the combination to individual application at least one Direct-Fed Microbials and phytase,The dosage wherein pressed more than about 1500FTU/kg feeds applies the phytase to the individual.

Description

Feed addictive composition

Technical field

The present invention relates to the groups at least one Direct-Fed Microbials and one or more high dose phytases Closing improves the method for fodder compound, and includes at least one Direct-Fed Microbials and one or more high dose inositols six The feed addictive composition of the combination of phosphatase.The invention further relates to purposes and kit.

Background technology

Necrotic enteritis is disease economically important in poultry species.It is because of pathogen C.perfringens (Clostridium perfringens) is infected and is generated, often with coccidiosis infection.In extreme circumstances, it leads to Gao Shui The flat death rate, and the adverse effect for leading to damage of intestines in subclinical levels and animal being showed.Mainly pass through feeding antibiotic Use control necrotic enteritis and its negative effect.But feeding antibiotic is used as the ban of growth promoter and is disappeared Expense person makes the producer be more difficult to prevent by C.perfringens the pressure used for reducing the antibiotic applied to meat producing animal And loss caused by relevant subclinical or clinical necrotic enteritis.Accordingly, there exist provide limitation or prevent by perfringens shuttle The chance of the solution of microbial damage.One this solution is with probiotics come the micro- life of sanatory enteron aisle Object fauna and the enterology ecosystem stablized.But just other are observed with the enzyme supplement of such as protease and carbohydrase Benefit.Although having contemplated that the combination of DFM and some enzymes, always without understanding the interaction between DFM and enzyme completely.This hair It is bright to be related to new specific combination, astoundingly significantly improve the production performance characteristic of animal.

Known nonruminant does not include endogenous phytase in stomach and small intestine or comprising the insignificant endogenous flesh of amount Alcohol six-phosphatase, therefore come in the alimentary canal of proximal end dependent on the plant and/or microorganism supplemented or fungi phytase Six phosphoric acid of hydrolysis of inositol (Pallauf, J. and Rimbach, G.Arch.Anim.Nutr., volume 1997,50,301-319 pages).It is logical Additional phytase is often added to the feed of nonruminant (such as poultry and pannage).Phytase be cereal and The major storage form of phosphorus in beans.By the effect of phytase, phytic acid, which usually hydrolyzes, generates rudimentary inositol Phosphoric acid and inorganic phosphate.

The present invention, which seeks to overcome, to be reduced with phytase in the animal with clinical or subclinical necrotic enteritis Beneficial effect correlation problem in some.

The present invention also seeks to overcome with phytic acid especially in the animal with clinical or subclinical necrotic enteritis In anti-nutritive properties correlation problem, lead to the validity for improving nutrients, minerals, vitamin and energy, and therefore change The biophysics feature of kind nonruminant.

Summary of the invention

The present invention initiative be the discovery that, the inventors discovered that, at least one DFM with high level (>1500FTU/kg is raised Material) phytase combination in there are synergistic effect beyong contemplation, have been displayed its by individual performance improve it is supreme In the horizontal level of the positive control with low-level C.perfringens and without necrotic enteritis.

The present inventor is by considering that such as Percentage mortality, weight gain and FCR demonstrate this discovery.These fronts Effect is entirely unexpected.

It is not intended to be limited to theory, inventors believe that, these effects can be by (such as being produced by pathogen compared with the control Gas capsular clostridium and/or Escherichia coli (Escherichia coli) attack) caused by the reduction of damage of intestines level cause.

The improvement of animal performance can be by preventing harmful bacteria colonizing in enteron aisle from being explained with pathology.In addition, Think that other beneficial effects of the present invention include that the hydrolysis of phytic acid increases, the ileum of protein and minerals is caused to disappear Rate improves.In addition to the benefit of the raising nutrients validity of individual, this may further result in less substrate and reaches hindgut use In bacterial fermentation so that the pathogen in enteron aisle is more difficult to colonize.The present invention's another benefit is that, and do not supplement phytic acid It is compared when enzyme, the generation of mucin is reduced.Accordingly, there exist the forfeitures of the endogenous of reduced levels, and same less substrate is used for Intestinal microflora is fermented.

Specifically, the present invention initiative be the discovery that at least one Direct-Fed Microbials (DFM) with high level (> 1500FTU/kg feeds) the combination of phytase there is significant beneficial effect to the performance of animal, including improve with It is one or more in lower：Feed conversion rate (FCR), ability (such as Nutrient digestibility, such as amino acid digestion for digesting raw material Rate), nitrogen reservation, survival rate, dressing percentage, growth rate, weightening, feed efficiency, animal is to the resistance of necrotic enteritis, individual Immune response.

Another surprising effect of the present invention is that it can reduce the excretion of the nutrients in muck (manure) and (such as subtract Nitrogen and phosphorus content in the muck of few individual).

In another aspect of this invention, such method, this method are provided：

I) it is used to improve the performance of individual；Or

Ii) it is used to improve the digestibility (such as Nutrient digestibility, such as amino acid digestibility) of the raw material in feed；Or

Iii it) is used to improve phosphorus (such as diet phosphorus) to absorb and retain；Or

Iv the effect of) being used to improve phytase；Or

V) it is used to improve nitrogen reservation；Or

Vi the resistance for) being used to improve individual to necrotic enteritis；Or

Vii it) is used to improve feed conversion rate (FCR)；Or

Viii it) is used to improve the weightening in individual；Or

Ix it) is used to improve the feed efficiency in individual；Or

X) it is used to adjust the immune response of (such as improvement) individual；Or

Xi) it is used to reduce the population of the pathogenic bacteria in individual gastrointestinal tract；Or

Xii the nutrients excretion in muck) is reduced；

This method includes the combination to individual application at least one Direct-Fed Microbials and phytase, wherein pressing Dosage more than about 1500FTU/kg feeds applies the phytase to the individual.

Another aspect of the present invention is the purposes of the combination of at least one Direct-Fed Microbials and phytase, In by more than about 1500FTU/kg feeds dosage use the phytase：

I) it is used to improve the performance of individual；Or

Iii it) is used to improve nitrogen reservation；Or

Iv it) is used to improve phosphorus (such as diet phosphorus) to absorb and retain；Or

V) the effect of being used to improve phytase；Or

Vii it) is used to improve feed conversion rate (FCR)；Or

Viii it) is used to improve the weightening in individual；Or

Ix it) is used to improve the feed efficiency in individual；Or

Xii the nutrients excretion in muck) is reduced.

The present invention also provides include at least one Direct-Fed Microbials and the addition of the feed of the combination of phytase Agent composition, the wherein phytase are by existing below in the feed addictive composition：

A. when being added in feed by least 50g/ tonnes of (MT) feed, by about 30,000FTU/g compositions or more；

B. when being added in feed by least 75g/ tonnes of (MT) feed, by about 20,000FTU/g compositions or more；

C. when being added in feed by least 100g/ tonnes of (MT) feed, by about 15,000FTU/g compositions or more；

D. when being added in feed by least 100g/ tonnes of (MT) feed, by about 15,000FTU/g compositions or more；

E. when being added in feed by least 150g/ tonnes of (MT) feed, by about 10,000FTU/g compositions or more；

F. when being added in feed by least 200g/ tonnes of (MT) feed, by about 7,500FTU/g compositions or more；

G. when being added in feed by least 300g/ tonnes of (MT) feed, by about 5,000FTU/g compositions or more；

And wherein the Direct-Fed Microbials are pressed from 2.5x10³CFU DFM:1FTU enzymes are to 6.7x10⁶CFU:The model of 1FTU enzymes It encloses and is present in the feed addictive composition.

In another aspect of this invention, the reagent of feed addictive composition and operation instructions comprising the present invention is provided Box.

In another aspect of this invention, the method for preparing feed addictive composition is provided comprising will be at least one straight It connects feeding microorganism to combine with phytase so that the dosage of the composition mysoinositol six-phosphatase：

A. it is about 30,000FTU/g compositions or more when being added in feed by least 50g/ tonnes of (MT) feed；

B. it is about 20,000FTU/g compositions or more when being added in feed by least 75g/ tonnes of (MT) feed；

C. it is about 15,000FTU/g compositions or more when being added in feed by least 100g/ tonnes of (MT) feed；

D. it is about 15,000FTU/g compositions or more when being added in feed by least 100g/ tonnes of (MT) feed；

E. it is about 10,000FTU/g compositions or more when being added in feed by least 150g/ tonnes of (MT) feed；

F. it is about 7,500FTU/g compositions or more when being added in feed by least 200g/ tonnes of (MT) feed；

G. it is about 5,000FTU/g compositions or more when being added in feed by least 300g/ tonnes of (MT) feed；

And in the feed addictive composition Direct-Fed Microbials dosage from 2.5x10³CFUDFM:1FTU enzymes To 6.7x10⁶CFU:In the range of 1FTU enzymes, and (optionally) packaging.

In another aspect of this invention, the feed of the feed addictive composition comprising the present invention is provided.

The present invention furthermore provides the method for preparing feed (feedstuff) comprising by feed ingredient and the present invention Feed addictive composition mixing.

On the other hand, the present invention is provided comprising feed addictive composition and at least one minerals and/or at least one The pre-composition of kind of vitamin, the feed addictive composition contain at least one Direct-Fed Microbials and phytase Combination, the wherein phytase are by existing below in the pre-composition：

And the Direct-Fed Microbials are pressed from 2.5x10³CFU DFM:1FTU enzymes are to 6.7x10⁶CFU:The range of 1FTU enzymes is deposited It is in the pre-composition.

The present invention also provides the present invention feed addictive composition or the present invention pre-composition or include the present invention feeding The feed of feed additives composition (or pre-composition), for preventing and/or treating globidiosis and/or gangrenosum acne intestines in individual It is scorching.

In a still further aspect thereof, the method that prevention is provided and/or treats necrotic enteritis and/or globidiosis, In to the feed addictive composition of a effective amount of present invention of individual application or the pre-composition of the present invention or include of the invention feeding The feed of feed additives composition (or pre-composition).

In the broad aspect of the present invention, method or purposes are furthermore provided：

I) it is used to improve the digestibility (such as Nutrient digestibility, such as amino acid digestibility) of the raw material in feed；Or

Ii) it is used to improve the performance of individual；Or

Iv the effect of) being used to improve phytase；Or

V) it is used to improve nitrogen reservation；Or

Vii it) is used to improve feed conversion rate (FCR)；Or

Viii it) is used to improve the weightening in individual；Or

Ix it) is used to improve the feed efficiency in individual；Or

Xii the nutrients excretion in muck) is reduced；

This method includes to individual administered with high dose phosphoric acid, together with the compound for reducing the pH in individual gastrointestinal tract (GIT). In this broad aspect, which can be suitably organic acid, essential oil or antibiotic.The organic acid can be selected from citric acid, prolong Fumarate, formic acid, propionic acid, lactic acid and benzoic acid.It is mould that the antibiotic can be selected from tetracycline, duomycin, terramycin, oleander Element, spiramvcin, Virginiamycin, Bacitracin Zinc, moenomycin (bambermycin), avilamycin and avoparcin.The essential oil (or plant extracts) Cinnemaldehyde, thymol, carvacrol, capsaicine, Eugenol (euganol) and alicin can be selected from.

Brief description

Fig. 1 displays supplement EnvivaPro and high phytic acid enzyme dosage (3000FTU/kg) in necrotic enteritis to attacking Hit the influence of the weight gain of broiler chicken (broiler) of the lower raising to 42 days.

Fig. 2 displays supplement EnvivaPro and high phytic acid enzyme dosage (3000FTU/kg) in necrotic enteritis to attacking Hit the influence of the FCR of broiler chicken of the lower raising to 42 days.

Fig. 3 shows that EnvivaPro and high phytic acid enzyme dosage (3000FTU/kg) cause necrotic enteritis attack The death rate influence.

Fig. 4 shows that EnvivaPro and high phytic acid enzyme dosage (3000FTU/kg) cause necrotic enteritis attack Damage of intestines severity influence.

Fig. 5 is shown under necrotic enteritis attack, in the case where existing and lacking Enviva Pro, the flesh of various dose The influence of FCR of the alcohol six-phosphatase to broiler chicken from the 0th day to the 21st day.

Fig. 6 shows the influence of FCR of the phytase of various dose to broiler chicken from the 6th day to the 35th day.

Fig. 7 is shown under necrotic enteritis attack, with and without Enviva Pro, various dose HiPhos^TMThe influence of FCR of the phytase to broiler chicken from the 0th day to the 21st day.

Fig. 8 is shown under necrotic enteritis attack, with and without Enviva Pro, different level HiPhos^TMPhytase to broiler chicken the 21st day shin bone (tibia) bone ash influence.

Detailed description of the invention

Preferably, it is exogenous for DFM for each in the enzyme of the present invention.In other words, preferably by enzyme It is added in DFM or is mixed with DFM.

Unless otherwise defined, all technical and scientific terms used herein all have disclosure fields common The normally understood meaning of technical staff.Singleton, etc. DICTIONARY OF MICROBIOLOGY AND MOLECULAR BIOLOGY, the 20th edition, John Wiley and Sons, New York (1994) and Hale＆Marham, THE HARPER COLLINS DICTIONARY OF BIOLOGY, Harper Perennial, NY (1991) provide for those skilled in the art The general dictionary of many terms used in the disclosure.

The disclosure is not limited to illustrative methods disclosed herein and material, it is any with those described herein method and The similar or equivalent method of material and material can be used in the implementation or test of the embodiment of the disclosure.Numberical range includes limit The number of the fixed range.Unless otherwise specified, being respectively, any nucleic acid sequence is write out with 5' to the directions 3' from left to right； Amino acid sequence is write out with amino to carboxyl direction from left to right.

Title provided herein is not these aspects or implementation to a variety of aspects of the disclosure or the limitation of embodiment Scheme can be obtained by considering the specification as a whole to refer to.Correspondingly, the term that will hereafter define will be by that will say Bright book refers to as a whole to be obtained more completely defining.

Amino acid is referred to amino acid name, trigram abbreviation or one-letter abbreviations herein.

The term as used herein " protein " includes protein, polypeptide and peptide.

The term as used herein " amino acid sequence " is synonymous with term " polypeptide " and/or term " protein ".In some feelings Under condition, term " amino acid sequence " is synonymous with term " peptide ".In some cases, term " amino acid sequence " and term " enzyme " are same Justice.

Term " protein " and " polypeptide " are used interchangeably herein.In the disclosure and claims, it can be used A conventional letter and three-letter codes for amino acid residue.3 alphanumeric codes of amino acid are in accordance with IUPACIUB biological chemical names The definition of joint committee (Joint Commission on Biochemical Nomenclature, JCBN).It should also be understood that Due to the degeneracy of genetic code, polypeptide can be nucleotide sequence coded by more than one.

Other definition of term can occur in entire this specification.Be more fully described exemplary implementation scheme it Before, it should be understood that the disclosure is not limited to described specific embodiment, because what these embodiments certainly can be changed.Also answer Understand, the term as used herein merely for description specific embodiment purpose, be not intended to it is in a limiting sense, because of this hair Bright range will be only limited by the following claims.

When providing numberical range, it should be appreciated that unless the context clearly dictates otherwise, also clearly disclose the range the upper limit and Each intermediate value between lower limit (to 1/10th of a position (unit) of lower limit).Arbitrary described value in the range or Each smaller range between other arbitrary described values or median in median and the range is also covered by the disclosure. These small range of upper and lower bounds can be independently include within the scope of this or exclude outside the range, and any of which, Neither one or two boundaries are included in each range in smaller range and are also covered by the disclosure, according in the range Depending on the arbitrary boundary clearly excluded.In the case where the range includes one or two of boundary, excludes these and wrapped The range of either one or two of the boundary included is also included in the disclosure.

It must be noted that unless the context clearly dictates otherwise, herein with singulative used in appended claims " one ", "one" and "the" include plural object.Thus, for example, it includes multiple this candidate substances to mention " enzyme " then, and mentions and " raising Material " includes then mentioning one or more feeds and their equivalent well known by persons skilled in the art, and so on.

The publication being discussed herein is intended merely to their disclosures before the submitting day of the application and provides.Herein Any content be all not interpreted as recognizing the prior art that these publications constitute this paper appended claims.

Enzyme for the present invention can be generated by solid culture or deep drainpipe, including batch processes, fed-batch side Method and continuous flow method.Culture complete in the medium, the culture medium include aqueous mineral salt culture medium, organic growth factors, Carbon and energy substance, the molecular oxygen and initial inoculation for also having one or more specific microbial species ready for use certainly Object.

Direct-Fed Microbials (DFM) for the present invention can be antipathogen Direct-Fed Microbials.It is used herein Term " antipathogen DFM " mean to inhibit (such as reduce or prevent) pathogenic microorganisms (such as C.perfringens and/or large intestine Bacillus and/or Salmonella ssp (Salmonella spp.) and/or campylobacter species (Campylobacter Spp.), preferably C.perfringens and/or Escherichia coli) the DFM that colonizes of enteral.

" the DFM measurement " hereafter instructed may be used to determine whether DFM is antipathogen DFM.

In one embodiment, inhibition bacterial strain (or disease-resistant original when selection " DFM measurement " assessment instructed herein Body DFM) as the DFM for the present invention.

Expediently, the DFM for the present invention can inhibit the one (or more) kinds in following pathogen：Clostridial species (Clostridium spp), such as C.perfringens and/or clostridium difficile (Clostridium difficile), and/or it is big Enterobacteria and/or Salmonella ssp and/or campylobacter species, at least one of preferably following pathogen (or it is more Kind)：Clostridial species, such as C.perfringens and/or clostridium difficile and/or Escherichia coli.

In one embodiment, the DFM for the present invention can inhibit the one (or more) kinds in following pathogen：Production Gas capsular clostridium and/or clostridium difficile and/or Escherichia coli, preferably C.perfringens and/or clostridium difficile, more preferable aerogenesis Capsular clostridium.

In one embodiment, individual can be one or more in such as following pathogen by following pathogen challenge：Clostridium Species, such as C.perfringens and/or clostridium difficile and/or Escherichia coli and/or Salmonella ssp and/or bending Bacillus species.

In one embodiment, individual can be attacked by C.perfringens and/or Escherichia coli.

DFM for the present invention is suitably bacterium living.

The present invention relates to high phytic acid enzyme dosages.

In general, giving phytase for example for broiler chicken and turkey by the range of about 500FTU/kg feeds, usually press The range of about 300FTU/kg feeds, which is given, is used for such as laying hen.

On the contrary, in the present invention, by least 1500FTU/kg feeds, suitably at least 2000FTU/kg feeds, suitably At least 3000FTU/kg feeds, suitably at least 5000FTU/kg feeds, such as 10000FTU/kg feeds it is horizontal by six phosphorus of inositol Sour enzyme is added in feed (or being added together with feed).

As will be understood by those skilled, the level of the phytase in feed addictive composition must be enough Allow by least 1500FTU/kg feeds, suitably at least 2000FTU/kg feeds, suitably at least 3000FTU/kg feeds, suitable Preferably at least phytase is added in feed by 5000FTU/kg feeds, the horizontal of such as 10000FTU/kg feeds.

It has been found by the present inventors that significant benefit can be obtained by the way that high phytic acid enzyme level is administered in combination with DFM. On the contrary, the dosage for individually increasing phytase does not generate notable benefit (for example, the nothing such as feed conversion rate significantly improves).Shape At striking contrast, increase phytase being combined at least one DFM (especially at least a kind of antipathogen DFM) When dosage, then notable and unexpected benefit is observed.This is entirely unexpected.

Expediently, the DFM for the present invention is the mixture of at least two DFM, suitably at least three kinds DFM.

In one embodiment, be used for the present invention DFM include from one or more of subordinate bacterium (or It is consisting essentially of, or be made from it)：Bacillus (Bacillus), enterococcus spp (Enterococcus), piece coccus Belong to (Pediococcus), saccharomyces (Saccharomyces), Bifidobacterium (Bifidobacterium), lactobacillus (Lactobacillus), lactococcus (Lactococcus), aspergillus (Aspergillus) and combinations thereof.

DFM for the present invention can include the bacterium from one or more of following species (or substantially by it Composition, or be made from it)：Bacillus subtilis (Bacillus subtilis), bacillus licheniformis (Bacillus Licheniformis), bacillus amyloliquefaciens (Bacillus amyloliquefaciens), Bacillus cercus (Bacillus cereus), enterococcus faecium (Enterococcus faecium), Pediococcus acidilactici (Pediococcus Acidilactici), saccharomyces cerevisiae (Saccharomyces cerevisiae), animal bifidobacteria animal subspecies (Bifidobacterium animalis spp animalis), lactobacillus reuteri (Lactobacillus reuteri), mouse Lee's sugar lactobacillus (Lactobacillus rhamnosus), Lactobacillus salivarius subspecies (Lactobacillus Salivarius ssp salivarius), Lactobacillus farciminis (Lactobacillus farciminis), Lactococcus lactis (Lactococcus lactis), clostridium butylicum (Clostridium butyricum), aspergillus oryzae (Aspergillus Oryzae) and combinations thereof.

DFM for the present invention can include that one or more of following bacterial strain is (or consisting essentially of, or by it Composition)：Bacillus subtilis BS18 (NRRL B-50633), bacillus subtilis BS278 (NRRL 50634), bacillus subtilis Bacterium 4-7d (NRRL B-50505), bacillus subtilis 3-5h (NRRL B-50507), bacillus subtilis AGTP BS3BP5 (NRRL B-50510), bacillus subtilis BS918 (NRRL B-50508), bacillus subtilis AGTP BS1013 (NRRL- 50509), bacillus subtilis AGTP 944 (NRRL B-50548), bacillus subtilis AGTP BS442 (NRRL B- 50542), bacillus subtilis AGTP BS1069 (NRRLB-50544), bacillus subtilis AGTP BS521 (NRRL B- 50545), bacillus subtilis BS2084 (NRRL B-50013), bacillus subtilis LSSA01 (NRRL B-50104), withered grass Bacillus B27 (NRRL B-50105), bacillus subtilis 3A-P4 (PTA-6506), bacillus subtilis 15A-P4 (PTA- 6507), bacillus subtilis 22C-P1 (PTA-6508), bacillus subtilis BL21 (NRRL B-50134), lichens gemma bar Bacterium BL21 (NRRL B-50134), bacillus licheniformis 3-12a (NRRL B-50504), bacillus licheniformis 4-2a (NRRL B- 50506), bacillus licheniformis 842 (NRRL B-50516), propionibacterium acide-propionici (Propionibacterium Acidipropionici) P261 (NRRL B-50131), propionibacterium acide-propionici P179 (NRRLB-50133), propionibacterium acide-propionici P169 (PTA 5271), propionibacterium acide-propionici P170 (PTA 5272), propionibacterium jensenii (Propionibacterium Jensenii) P63 (NRRL B-30979), propionibacterium jensenii P195 (NRRL B-50132), Lactococcus lactis ID7 (PTA 6103), Lactococcus lactis JD19 (PTA 6104), lactobacillus acidophilus (Lactobacillus acidophilus) A2020 (NRRL B-30977), lactobacillus acidophilus A4000h (NRRL B-30978), lactobacillus acidophilus PIBc6 (NRRL B-50103), Lactobacillus brevis (Lactobacillus brevis) LBR 1000 (NRRL B-30982), Lactobacillus casei (Lactobacillus Casei) LC222 (NRRL B-30983), Yue Shi lactobacillus (Lactobacillus johnsonii) PLCB6 (NRRL B- 50518), Lactobacillus salivarius (Lactobacillus salivarius) o246i33w (NRRL B-50102), Lactobacillus brevis AJ25 (PTA-6099), Lactobacillus brevis HE17 (PTA-6100), Lactobacillus brevis 1E-1 (PTA-6509), lactobacillus lactis (Lactobacillus lactis) Cl15 (PTA-6101), lactobacillus lactis DJ6 (PTA-6102), Lactobacillus rhamnosus (CNCM I-3698), Lactobacillus farciminis (CNCM I-3699), enterococcus faecium EF141 (EN-1) (NRRL B-30981), dung intestines Coccus 2-1d (NRRL B-50519), Pediococcus acidilactici PIJe3 (NRRL B-50101), Pediococcus acidilactici o246e42 (NRRL B-50171) and combinations thereof.

DFM for the present invention can include more than one, suitably two or more, suitably at least three kinds of bacillus Species (Bacillus spp).

DFM for the present invention can include that one or more of following bacterial strain is (or consisting essentially of, or by it Composition)：Bacillus subtilis strain 3A-P4 (PTA-6506), 15A-P4 (PTA-6507), 22C-P1 (PTA-6508), 2084 (NRRL B-500130), LSSA01 (NRRL-B-50104), BS27 (NRRL B-50105), BS 18 (NRRL B-50633) and BS 278 (NRRL B-50634) or combinations thereof.

In one embodiment, the DFM for being used for the present invention can include that one or more of following bacterial strain is (or basic On be made from it, or be made from it)：Bacillus subtilis strain 15A-P4 (PTA-6507), 2084 (NRRL B-500130), LSSA01(NRRL-B-50104)。

In one embodiment, the phytase for the present invention is six phosphorus of 6-phytase or 3- inositols Sour enzyme, preferably 6-phytase.

In one embodiment, the phytase for the present invention can be selected from：Escherichia coli phytic acid Enzyme or Buttiauxella (Buttiauxella) phytase or six phosphorus of Citrobacter (Citrobacter) inositol Sour enzyme or Hafnia (Hafnia) phytase or aspergillus phytase or Penicillium (Penicillium) phytase or trichoderma (Trichoderma) phytase or Escherichia coli inositol six Phosphatase or Hansenula (Hansenula) phytase, or combinations thereof.

In one embodiment, the phytase for the present invention can be one or more in following 6-phytase：Escherichia coli, Buttiauxella species (Buttiauxella spp) or citric acid bacterium species (Citrobacter spp), such as Bu Shi citric acid bacteriums (Citrobacter braakii), including a combination thereof.

Expediently, the phytase for the present invention can be one or more PhyzymeXP^TM、AxtraPhy^TM, Or Ronozyme HiPhos^TMOr Quantum^TMOr Quantum Blue^TM。

Expediently, phytase can be more than one phytases, such as at least two kinds of or at least three kinds of inositols The mixture of six-phosphatase.

In one embodiment, it is preferred to ground, the phytase (example active at a low ph for the present invention Such as, active between pH2 and pH5.5).

In one embodiment, it is preferred to ground, the phytase for the present invention has the optimal pH (example under low pH Such as, with the optimal pH in the range of pH2 and pH5.5).

In one embodiment, there is clinical or subclinical enteropathy attack in individual.

In one embodiment, the attack of clinical or subclinical enteropathy can be by arbitrary pathogen or arbitrary pathogenic bacteria (such as C.perfringens or Escherichia coli) cause.

In one embodiment, in feed addictive composition Direct-Fed Microbials dosage can from 3.8x10³CFU DFM:1FTU enzymes are to 2.0x10⁵CFU:In the range of 1FTU enzymes.

Direct-Fed Microbials (DFM)

Term " microorganism (microbial) " herein can be used interchangeably with " microbial body (microorganism) ".

Preferably, DFM includes viable microbial.Preferably, DFM includes bacterium living or viable yeast or fungi living.

Preferably, DFM includes bacterium living.

Term " viable microbial " means the microorganism that has metabolic activity or can break up.

In one embodiment, DFM can be spore forming bacteria, therefore term DFM can be by spore, such as bacterium spore Son composition, or contain spore, such as bacterial spore.Therefore, in one embodiment, the term as used herein " viable microbial " May include microbial spore, such as endospore or conidium.

In another embodiment, the DFM in feed addictive composition of the invention is by microbial spore, such as Endospore or conidium composition, or do not contain microbial spore, such as endospore or conidium.

Microorganism can be naturally occurring microorganism or its can be microbial.Microorganism can also be suitable micro- The combination of biology.

In some respects, DFM of the invention can be it is following in it is one or more：Bacterium, yeast, fungi.

Preferably, DFM of the invention is probiotic microorganisms.

In the present invention, term Direct-Fed Microbials (DFM) cover Direct-fed bacterium, Direct-fed yeast, directly Feed fungi and combinations thereof.

Preferably, DFM is Direct-fed bacterium.

Preferably, DFM is the combination for including two or more bacteriums (such as three kinds or a variety of or four kinds or a variety of)；Or Person DFM is the combination for including two or more bacterium bacterial strains (such as three kinds or a variety of or four kinds or a variety of).

Preferably, bacterium is separation.

Expediently, DFM can include from the bacterium of one or more of subordinate：Lactobacillus, lactococcus, chain Coccus (Streptococcus), bacillus, Pediococcus, enterococcus spp, Leuconostoc (Leuconostoc), meat Eat Bacillus (Carnobacterium), Propionibacterium (Propionibacterium), Bifidobacterium, fusobacterium and huge Spherical Pseudomonas (Megasphaera) and combinations thereof.

In one embodiment, DFM can be selected from following Bacillus spec：Bacillus subtilis, wax-like gemma Bacillus, bacillus licheniformis, bacillus amyloliquefaciens and bacillus pumilus (Bacillus pumilus).

In one embodiment, DFM can be Bacillus strain.

In one embodiment, DFM can be selected from：

Bacillus subtilis BS18 (NRRL B-50633), bacillus subtilis BS278 (NRRL50634), withered grass gemma Bacillus 4-7d (NRRL B-50505), bacillus subtilis 3-5h (NRRL B-50507), bacillus subtilis AGTP BS3BP5 (NRRL B-50510), bacillus subtilis BS918 (NRRL B-50508), bacillus subtilis AGTP BS1013 (NRRL- 50509), bacillus subtilis AGTP 944 (NRRL B-50548), bacillus subtilis AGTP BS442 (NRRL B- 50542), bacillus subtilis AGTP BS1069 (NRRL B-50544), bacillus subtilis AGTP BS521 (NRRL B- 50545), bacillus subtilis BS2084 (NRRL B-50013), bacillus subtilis LSSA01 (NRRL B-50104), withered grass Bacillus B27 (NRRL B-50105), bacillus subtilis 3A-P4 (PTA-6506), bacillus subtilis 15A-P4 (PTA- 6507), bacillus subtilis 22C-P1 (PTA-6508), bacillus subtilis BL21 (NRRL B-50134) and combinations thereof.

To avoid generating query, bacillus subtilis LSSA01 (NRRL B-50104) is sometimes referred to as bacterial strain BS8.

In one embodiment, DFM can be selected from：

Bacillus subtilis strain AGTP BS 1068 (NRRL B-50543) or bacillus pumilus KX11-1 (NRRL B-50546) and combinations thereof.

In one embodiment, DFM is selected from：

Bacillus licheniformis BL21 (NRRL B-50134), bacillus licheniformis 3-12a (NRRL B-50504), lichens bud Spore bacillus 4-2a (NRRL B-50506) and bacillus licheniformis 842 (NRRL B-50516).

In one embodiment, DFM can be the combination for including two or more Bacillus strains.

In one embodiment, DFM can be two or more bacillus subtilis strains 3A-P4 (PTA-6506), 15A-P4(PTA-6507)、22C-P1(PTA-6508)、2084(NRRL B-500130)、LSSA01(NRRL-B-50104)、 The combination of BS27 (NRRL B-50105), BS 18 (NRRL B-50633) and BS 278 (NRRL B-50634).

In preferred embodiments, DFM can include that one of following combination is (or consisting essentially of, or by its group At)：

Bacillus subtilis LSSA01 (NRRL B-50104)+bacillus subtilis BS18 (NRRL B-50633)+withered grass Bacillus 22C-P1 (PTA-6508)；Or

Bacillus subtilis LSSA01 (NRRL B-50104)+bacillus subtilis 3A-P4 (PTA-6506)+withered grass gemma Bacillus BS2084 (NRRL B-50013)；Or

Bacillus subtilis LSSA01 (NRRL B-50104)+bacillus subtilis 3A-P4 (PTA-6506)+withered grass gemma Bacillus BS2084 (NRRL B-50013)+bacillus subtilis 15A-P4 (PTA-6507)；Or

Bacillus subtilis 3A-P4 (PTA-6506)+bacillus subtilis 22C-P1 (PTA-6508)+bacillus subtilis BS2084(NRRL B-50013)；Or

Bacillus subtilis LSSA01 (NRRL B-50104)+bacillus subtilis 15A-P4 (PTA-6507)+withered grass bud Spore bacillus BS2084 (NRRL B-50013)；Or

Bacillus subtilis 3A-P4 (PTA-6506)+bacillus subtilis BS18 (NRRL B-50633)+bacillus subtilis Bacterium 22C-P1 (PTA-6508)；Or

Bacillus subtilis 3A-P4 (PTA-6506)+bacillus subtilis BS2084 (NRRL B-50013)+withered grass gemma Bacillus BS278 (NRRL 50634).

Bacterial strain 3A-P4 (PTA-6506), 15A-P4 (PTA-6507) and 22C-P1 (PTA-6508) can be trained from U.S. typical case Support the open acquisition of object collection (American Type Culture Collection, ATCC).

Bacterial strain 2084 (NRRL B-500130), LSSA01 (NRRL-B-50104), BS27 (NRRL B-50105) can be from U.S.s Agricultural research institute of state collection (Agricultural Research Service Culture Collection, NRRL) It is open to obtain.Strains B. subtilis LSSA01 is sometimes referred to as bacillus subtilis 8.

These bacterial strains are instructed in US 7,754,469B2.

Danisco u s company (Danisco USA Inc.) (W227N752Westmound Dr.Waukesha, WI 53186, USA) following preserved material is preserved in american agriculture research aircraft according further to budapest treaty (Budapest Treaty) Structure collection (1815North University Street, Peoria, Illinois 61604, United States of America), the preservation date and preserving number of original deposit is detailed below：

Bacillus licheniformis 3-12a	NRRL B-50504	On May 13rd, 2011
			Bacillus licheniformis 4-2a	NRRL B-50506	On May 13rd, 2011
Bacillus licheniformis BL842	NRRL B-50516	On May 20th, 2011
			Bacillus pumilus AGTP KXII-1	NRRL B-50546	On August 5th, 2011
Bacillus pumilus AGTP BS1068	NRRL B-50543	On August 4th, 2011
			Bacillus subtilis BS18	NRRL B-50633	On January 9th, 2012
Bacillus subtilis BS278	NRRL B-50634	On January 9th, 2012
			Bacillus subtilis 4-7d	NRRL B-50505	On May 13rd, 2011
Bacillus subtilis 3-5h	NRRL B-50507	On May 13rd, 2011
			Bacillus subtilis AGTP BS3BP5	NRRL B-50510	On May 13rd, 2011
Bacillus subtilis AGTP BS918	NRRL B-50508	On May 13rd, 2011
			Bacillus subtilis AGTP BS1013	NRRL B-50509	On May 13rd, 2011
Bacillus subtilis AGTP 944	NRRL B-50548	On August 11st, 2011
			Bacillus subtilis AGTP BS442	NRRL B-50542	On August 4th, 2011
Bacillus subtilis AGTP BS1069	NRRL B-50544	On August 4th, 2011
			Bacillus subtilis AGTP BS521	NRRL B-50545	On August 4th, 2011

Danisco u s company (Danisco USA Inc.) (W227N752Westmound doctors Waukesha, WI 53186, the U.S.) authorize DuPont Nutrition BioSciences Co., Ltd. (DuPont Nutrition Biosciences ApS) (Langebrogade 1, PO Box 17, DK-1001, Copenhagen K, Denmark) refers to these guarantors in the present patent application The biomaterial of Tibetan, and agree to the sky and that irrevocablely the public can get the material of institute preservation.

In some embodiments, DFM can be the combination for including the bacillus subtilis strain being described in detail in following table.

In one embodiment, DFM can be selected from lactococcus species (Lactococcus spp).

In one embodiment, DFM can be selected from one or more in following lactococcus species：Butterfat galactococcus (Lactococcus cremoris) and Lactococcus lactis and combinations thereof.

In one embodiment, DFM can be selected from following bacterial strain or combinations thereof：

Lactococcus lactis ID7	PTA 6103
		Lactococcus lactis JD19	PTA 6104

In one embodiment, DFM can be Lactobacillus species (Lactobacillus spp).

In one embodiment, DFM can be selected from following Lactobacillus species：Lactobacillus buchneri (Lactobacillus Buchneri), lactobacillus acidophilus, Lactobacillus casei, Kefir grains lactobacillus (Lactobacillus kefiri), double qi lactobacillus (Lactobacillus bifidus), Lactobacillus brevis, Lactobacillus helveticus (Lactobacillus helveticus), class cheese Lactobacillus (Lactobacillus paracasei), Lactobacillus rhamnosus, Lactobacillus salivarius, lactobacillus curvatus (Lactobacillus curvatus), lactobacillus bulgaricus (Lactobacillus bulgaricus), Lactobacillus saki (Lactobacillus sakei), lactobacillus reuteri, lactobacillus fermenti (Lactobacillus fermentum), sausage breast bar Bacterium, lactobacillus lactis, Lactobacillus delbrueckii (Lactobacillus delbreuckii), lactobacillus plantarum (Lactobacillus Plantarum), class lactobacillus plantarum (Lactobacillus paraplantarum), Lactobacillus farciminis, Lactobacillus rhamnosus, Lactobacillus crispatus (Lactobacillus crispatus), Lactobacillus gasseri (Lactobacillus gasseri), Yue Shi breasts Bacillus and Lactobacillus Jensenii (Lactobacillus jensenii) and its arbitrary combination.

In one embodiment, DFM is selected from following bacterial strain or combinations thereof：

Lactobacillus acidophilus A2020	NRRL B-30977
		Lactobacillus acidophilus A4000h	NRRL B-30978
Lactobacillus acidophilus PIBc6	NRRL B-50103
		Lactobacillus brevis LBR 1000	NRRL B-30982
Lactobacillus casei LC222	NRRL B-30983
		Yue Shi lactobacillus PLCB6	NRRL B-50518
Lactobacillus salivarius o246i33w	NRRL B-50102
		Lactobacillus brevis AJ25	PTA-6099
Lactobacillus brevis HE17	PTA-6100
		Lactobacillus brevis 1E-1	PTA-6509
Lactobacillus lactis Cl15	PTA-6101
		Lactobacillus lactis DJ6	PTA-6102
Lactobacillus rhamnosus	CNCM I-3698
		Lactobacillus farciminis	CNCM I-3699

In one embodiment, DFM can be selected from one or more of following bacterial strain：Lactobacillus rhamnosus CNCM- I-3698 and Lactobacillus farciminis CNCM-I-3699.These bacterial strains by Sorbial (Route de Spay 72700Allonnes, France) French Organism Depositary (Collection Nationale de Cultures are deposited on December 8th, 2006 De Microorganims) (CNCM) (25, Rue due Docteur Roux, F75724Paris Cedex 15, France), it protects Hide object all authority, ownership and interests be subsequently transferred to Danisco France SAS (20, Rue de Brunel, 75017Paris, France).

Danisco France SAS authorize DuPont Nutrition BioSciences Co., Ltd. (Langebrogade1, PO Box 17, DK-1001, Copenhagen K, Denmark) in the present patent application with reference to the biomaterial of these preservations, and without reserve Ground and irrevocable the material public for agreeing to institute's preservation can get.

In one embodiment, DFM can be can be from Danisco Animal nutrition company (Danisco Animal Nutrition it) obtainsProduct.

In one embodiment, DFM can come from Pediococcus.

In one embodiment, DFM can come from following Pediococcus species：Pediococcus acidilactici, such as following lactic acid sheet It is one or more in meningitidis strains PIJe3 (NRRL B-50101) or o246e42 (NRRL B-50171).

In one embodiment, DFM can be selected from following Bifidobacteria (Bifidobacteria spp)：Breast Bifidobacterium (Bifidobacterium lactis), bifidobacterium bifidum (Bifidobacterium bifidium), length are double Discrimination bacillus (Bifidobacterium longum), B. animais (Bifidobacterium animalis), short bifid Bacillus (Bifidobacterium breve), bifidobacterium infantis (Bifidobacterium infantis), chain bifid bar Bacterium (Bifidobacterium catenulatum), false chain Bifidobacterium (Bifidobacterium Pseudocatenulatum), bifidobacterium adolescentis (Bifidobacterium adolescentis) and angle Bifidobacterium (Bifidobacterium angulatum) and its arbitrary combination.

In one embodiment, DFM can be Enterococcus species (Enterococcus spp).

In one embodiment, DFM can come from enterococcus faecium species.

In one embodiment, DFM can be selected from enterococcus faecium EF141 (EN-1) (NRRL B-30981), dung intestines ball Bacterium 2-1d (NRRL B-50519) and combinations thereof.

In one embodiment, DFM can come from Propionibacterium.

In one embodiment, DFM can be selected from following Propionibacterium species or combinations thereof：Propionibacterium acide-propionici and Propionibacterium jensenii.

In one embodiment, DFM can be selected from following Propionibacterium species or combinations thereof：

Propionibacterium acide-propionici P261	NRRL B-50131
		Propionibacterium acide-propionici P179	NRRL B-50133
Propionibacterium acide-propionici P169	PTA 5271
		Propionibacterium acide-propionici P170	PTA 5272
Propionibacterium jensenii P63	NRRL B-30979
		Propionibacterium jensenii P195	NRRL B-50132

Expediently, DFM can include the bacterium from one or more of following species：Bacillus subtilis, lichens Bacillus, bacillus amyloliquefaciens, enterococcus faecium, Enterococcus species and Pediococcus species (Pediococcus spp), It is Lactobacillus species, Bifidobacteria, lactobacillus acidophilus, Pediococcus acidilactici, Lactococcus lactis, bifidobacterium bifidum, withered Careless bacillus, Te Shi Propionibacteriums (Propionibacterium thoenii), Lactobacillus farciminis, Lactobacillus rhamnosus, angstrom Family name's Megasphaera (Megasphaera elsdenii), clostridium butylicum, B. animais animal subspecies, lactobacillus reuteri, wax Shape bacillus, Lactobacillus salivarius subspecies, Propionibacterium species (Propionibacteria sp) and combinations thereof.

Direct-fed bacterium for the present invention may belong to same type (genus and species and bacterial strain) or can include genus and species And/or the mixture of bacterial strain.

Expediently, the DFM of the present invention can be product or the microorganism being contained in those products those of in following table In it is one or more：

In one embodiment, expediently, DFM can be

It can be bought from Danisco A/S BJ Rep Office, be 2084 preserving number NRRl B- of Bacillus strain 50013, Bacillus strain LSSAO1 preserving numbers NRRL B-50104 and Bacillus strain 15A-P4ATCC preserving numbers The combination (as taught in US 7,754,469B --- be hereby incorporated by reference) of PTA-6507.

In one embodiment, DFM can be bacillus licheniformis BL21 (NRRL B-50134).

AgTech Products, Inc. (W227N752Westmound Drive, Waukesha, WI53186, the U.S.) are pressed Following biological deposit is preserved in american agriculture research institution collection (NRRL) according to budapest treaty, original is detailed below The preservation date and preserving number of beginning preserved material：

Bacillus licheniformis BL21	NRRL B-50134	On April 15th, 2008
			Enterococcus faecium EF141 (EN-1)	NRRL B-30981	On October 30th, 2006
Enterococcus faecium 2-1d	NRRL B-50519	On June 3rd, 2011
			Lactobacillus acidophilus A2020	NRRL B-30977	On October 24th, 2006
Lactobacillus acidophilus A4000h	NRRL B-30978	On October 24th, 2006
			Lactobacillus acidophilus PIBc6	NRRL B-50103	On January 18th, 2008
Lactobacillus brevis LBR 1000	NRRL B-30982	On October 30th, 2006
			Lactobacillus casei LC222	NRRL B-30983	On October 30th, 2006
Yue Shi lactobacillus PLCB6	NRRL B-50518	On June 3rd, 2011
			Lactobacillus salivarius o246i33w	NRRL B-50102	On January 18th, 2008
Pediococcus acidilactici PIJe3	NRRL B-50101	On January 18th, 2008
			Pediococcus acidilactici o246e42	NRRL B-50171	On August 29th, 2008

AgTech Products, Inc authorize DuPont Nutrition BioSciences Co., Ltd. (Langebrogade 1, PO Box 17, DK-1001, Copenhagen K, Denmark) in the present patent application with reference to the biomaterial of these preservations, and without reserve Ground and irrevocable the material public for agreeing to institute's preservation can get.

AgTech Products, Inc. (W227N752Westmound Drive, Waukesha, WI53186, the U.S.) are also According to budapest treaty by following biological deposit be preserved in American type culture collection (ATCC) (Manassas, VA 20110-2209, the U.S.) (PTA references) or american agriculture research institution collection (NRRL references), it is detailed below original The preservation date and preserving number of preserved material --- these bacterial strains are mentioned in listed patent in the table, therefore are open can get Bacterial strain：

In one embodiment, the DFM for the present invention is selected from：Bacillus subtilis BS18 (NRRL B-50633), Bacillus subtilis BS278 (NRRL 50634), bacillus subtilis 4-7d (NRRL B-50505), bacillus subtilis 3-5h (NRRL B-50507), bacillus subtilis AGTP BS3BP5 (NRRL B-50510), bacillus subtilis BS918 (NRRL B-50508), bacillus subtilis AGTP BS1013 (NRRL-50509), (the NRRL B- of bacillus subtilis AGTP 944 50548), bacillus subtilis AGTP BS442 (NRRL B-50542), bacillus subtilis AGTP BS1069 (NRRL B- 50544), bacillus subtilis AGTP BS521 (NRRL B-50545), bacillus subtilis BS2084 (NRRL B-50013), Bacillus subtilis LSSA01 (NRRL B-50104), bacillus subtilis B 27 (NRRL B-50105), bacillus subtilis It is 3A-P4 (PTA-6506), bacillus subtilis 15A-P4 (PTA-6507), bacillus subtilis 22C-P1 (PTA-6508), withered Careless bacillus BL21 (NRRL B-50134), bacillus licheniformis BL21 (NRRL B-50134), bacillus licheniformis 3-12a (NRRL B-50504), bacillus licheniformis 4-2a (NRRL B-50506), bacillus licheniformis 842 (NRRL B-50516), Propionibacterium acide-propionici P261 (NRRL B-50131), propionibacterium acide-propionici P179 (NRRL B-50133), propionibacterium acide-propionici P169 (PTA 5271), propionibacterium acide-propionici P170 (PTA 5272), propionibacterium jensenii P63 (NRRL B-30979), Zhan Shi Propionibacterium P195 (NRRL B-50132), Lactococcus lactis ID7 (PTA 6103), Lactococcus lactis JD19 (PTA 6104), Lactobacillus acidophilus A2020 (NRRL B-30977), lactobacillus acidophilus A4000h (NRRL B-30978), lactobacillus acidophilus PIBc6 (NRRL B-50103), Lactobacillus brevis LBR 1000 (NRRL B-30982), Lactobacillus casei LC222 (NRRL B-30983), Yue Shi lactobacillus PLCB6 (NRRL B-50518), Lactobacillus salivarius o246i33w (NRRL B-50102), Lactobacillus brevis AJ25 (PTA-6099), Lactobacillus brevis HE17 (PTA-6100), Lactobacillus brevis 1E-1 (PTA-6509), lactobacillus lactis Cl15 (PTA- 6101), lactobacillus lactis DJ6 (PTA-6102), Lactobacillus rhamnosus (CNCM I-3698), Lactobacillus farciminis (CNCM I- 3699), enterococcus faecium EF141 (EN-1) (NRRL B-30981), enterococcus faecium 2-1d (NRRL B-50519), Pediococcus acidilactici PIJe3 (NRRL B-50101), Pediococcus acidilactici o246e42 (NRRL B-50171) and combinations thereof.

Expediently, DFM can include the yeast from Blastocystis species (Saccharomyces spp).

Preferably, wait for that DFM is it has been generally acknowledged that safety and the microorganism preferably ratified through GRAS used according to the present invention.

Those skilled in the art will readily recognize that from for food and/or agricultural and it has been generally acknowledged that suitable for animal food The specific species and/or bacterial strain of microorganism in category described herein.

Preferably, DFM used according to the invention is adapted for the DFM of animal edible.

Advantageously, if product is feed or feed addictive composition, DFM living should provide feed or feeding in retailer Feed additives composition keeps effective for normal " the guaranteeing the quality " of the entire product of sale or during " effective " date.It is desired Time span and normal shelf life will be different because of feed difference, and those skilled in the art will recognize that shelf life will The difference depending on feed type, feed size, storage temperature, treatment conditions, packaging material and packaging facilities.

In some embodiments, it is important that DFM is resistant to be heated, and is heat-resisting.By feed pill (pelleted) especially true in the case of.Therefore, in one embodiment, DFM can be themoduric, such as heat-resisting Bacterium, including such as Bacillus spec.

In some embodiments, it may be preferred that DFM is spore-bearing bacterium, such as bacillus, such as bacillus Species.When growth conditions is unfavorable, bacillus can form stable endospore, and it to heat, pH, moisture and disappears Toxic agent has extremely strong resistance.

In one embodiment, expediently, DFM can reduce or prevent pathogenic microorganisms (such as C.perfringens and/or Escherichia coli and/or Salmonella ssp and/or campylobacter species, preferably C.perfringens and/or Escherichia coli) It colonizes in the intestine.

The DFM of the present invention can be the DFM of any suitable.In one embodiment, " DFM surveys can be measured with following It is fixed " determine that microorganism becomes the adaptability of DFM.To avoid generating query, in one embodiment, according to teaching herein " DFM measurement " be selected as inhibition bacterial strain (or antipathogen DFM) DFM be suitable for the present invention (i.e. according to the present invention Feed addictive composition) DFM.

DFM is measured：

Pipe is inoculated with by the representative pathogens body from representative cluster.

Into the pipe being inoculated with, addition is from a manner of aerobic or the supernatant of the potential DFM of anaerobism mode culture, and is incubated.

After incubation, for the optical density (OD) of each cause of disease bulk measurement control tube and supernatant processing tube.

The bacterium colony for generating (potential DFM) bacterial strain for reducing OD compared with the control is classified as inhibition bacterial strain (or disease-resistant original Body DFM).

DFM used herein is illustrated in more detail in US2009/0280090 (being hereby incorporated by reference) to measure.

Preferably, it is one of the following (or multiple) for the representative pathogens body of measurement：Fusobacterium, such as perfringens Clostridium and/or clostridium difficile and/or Escherichia coli and/or Salmonella ssp and/or campylobacter species.At one In preferred embodiment, the measurement is carried out with one or more in following：C.perfringens and/or clostridium difficile and/or Escherichia coli, preferably C.perfringens and/or clostridium difficile, more preferable C.perfringens.

In one embodiment, DFM of the invention is preferably antipathogen.

The term as used herein " antipathogen " means that DFM offsets the effect (such as negative effect) of pathogen.

In one embodiment, for determine DFM whether be the present invention antipathogen, can be measured using above-mentioned DFM. If DFM is classified as inhibition bacterial strain in above-mentioned " DFM measurement ", especially when pathogen is C.perfringens, then DFM is considered as antipathogen or antipathogen DFM.

In one embodiment, antipathogen DFM can be one or more in following bacterium：

Bacillus subtilis strain 2084, preserving number NRRl B-50013,

Bacillus subtilis strain LSSAO1, preserving number NRRL B-50104；

Bacillus subtilis strain 15A-P4, ATCC preserving number PTA-6507；

Bacillus subtilis 3A-P4, ATCC preserving number PTA-6506；With

Bacillus subtilis strain BS27, ATCC preserving number NRRL B-50105.

To avoid generating query, these bacterial strains are obtainable, and are referred in US 7,754,459B.

In one embodiment, DFM used according to the invention is not the preserving number taught in WO2008/016214 For 17 bacterial strains of Lactobacillus gasseri BNR of KCTC 10902BP.

Preferably, DFM is not inactivation microorganism.

In one embodiment, it is the group for including one or more DFM microorganisms as described herein used herein of DFM Close object.The composition can additionally comprise the enzyme of the present invention.

It can be fed the composition as Direct-Fed Microbials (DFM) to animal.

One or more carriers or other compositions can be added into DFM.

DFM can be presented with various physical forms, for example, in the form of surface charging (top dress), it is for animals for use as liquid Draught or to be added to water-soluble concentrate, gelatine capsule or the gel form in newborn substitute.

In an embodiment of surface dosing form, by the tunning of freeze-drying be added to carrier (such as breast Clearly, maltodextrin, sucrose, dextrose, lime stone (calcium carbonate), rice husk, yeast culture, dry starch and/or sodium silicoaluminate) In.

It, will in for an embodiment of liquid drencs or the water-soluble concentrate of newborn substitute replenishers The tunning of freeze-drying is added to water-solubility carrier (such as whey, maltodextrin, sucrose, dextrose, dry starch, silicon Sodium aluminate) in, and adding liquid is to form drencs or into breast or newborn substitute add replenishers.

In an embodiment of hard gelatin capsule forms, by the tunning of freeze-drying be added to carrier (such as whey, Maltodextrin, sugar, lime stone (calcium carbonate), rice husk, yeast culture, dry starch and/or sodium silicoaluminate) in.

In one embodiment, bacterium and carrier are closed in degradable gelatine capsule.At one of gel form In embodiment, the tunning of freeze-drying is added to carrier (such as vegetable oil, sucrose, silica, polysorbate 80, propylene glycol, butylated hydroxyanisol, citric acid, ethoxyquin (ethoxyquin) and/or artificial tanning agent) in formed Gel.

DFM can (include but not limited to optionally growth substrate, enzyme, sugar, carbohydrate, extract and growth promotion with additive Micro- ingredient) drying agent mixing.Sugar may include following：Lactose, maltose, dextrose, maltodextrin, glucose, fruit Sugar, mannose, Tagatose (tagatose), sorbose, gossypose and galactolipin.Alone or in combination form sugar ranging from 50-95%.Extract may include yeast or dry yeast fermentation solubles, ranging from 5-50%.Growth substrate may include：Pancreas egg White casein hydrolysis, ranging from 5-25%；Sodium lactate, ranging from 5-30%；And Tween 80, ranging from 1-5%.Carbohydrate can Including mannitol, sorbierite, adonite and arabitol.The ranging from 5-50% of the carbohydrate of form alone or in combination.It is micro- at Point it may include following：Calcium carbonate, ranging from 0.5-5.0%；Calcium chloride, ranging from 0.5-5.0%；Dipotassium hydrogen phosphate, ranging from 0.5-5.0%；Calcium phosphate, ranging from 0.5-5.0%；Albumen manganese, ranging from 0.25-1.00%；And manganese, ranging from 0.25- 1.0%.

It, can be by one or more cultures and one or more carriers (if if use) to prepare DFM as described herein It is added to belt or arm mixer, and is mixed about 15 minutes, but the time can be increased or decreased.Each component is mixed so that producing The homogeneous mixture of raw culture and carrier.The preferably dry flowable powder of final product.Then by DFM or DFM can be included Composition be added in animal feed or feed premix, be added in the water of animal, or with it is known in the art other Mode application (is preferably administered simultaneously with the enzyme of the present invention).One or more DFM or as described herein groups as described herein can be used It closes object and supplements animal feed.

" mixtures of at least two bacterial strains " mean two kinds, three kinds, four kinds, five kinds, six kinds or even more a variety of bacterial strains Mixture.In some embodiments of the mixture of bacterial strain, ratio can be changed to 99% from 1%.The mixture of bacterial strain its His embodiment is 25% to 75%.The other embodiments of the mixture of bacterial strain are that each bacterial strain is about 50%.Mixed When closing object comprising two or more bacterial strains, ratio or be present in mixture in varing proportions that bacterial strain can be substantially equal.

In one embodiment, DFM can be the alive or dead microorganism used in the form of detaching or is semi-detached. DFM can be combined with the growth medium for cultivating it wherein or not combined.

In one embodiment, when cultivating on appropriate media, DFM can generate Colony Forming Unit.This is appropriate Medium can include feed or feed ingredient (or being made from it).

In one embodiment, when cultivating on appropriate media, DFM cannot generate Colony Forming Unit.This is suitably situated between Matter can include feed or feed ingredient (or being made from it).

Whether can or can not generate that Colony Forming Unit --- cell still may be used when no matter DFM is cultivated on appropriate media With with metabolic activity (for example, even if they cannot be separated).

In one embodiment, DFM can be used as dead cell to apply.

In one embodiment, DFM can be used as viable microbial to apply.

DFM can be properly joined into.

Expediently, the dosage of DFM can be between about 1 × 10 in feed³CFU/g feeds are to about 1 × 10⁹Between CFU/g feeds, Suitably between about 1 × 10⁴CFU/g feeds are to about 1 × 10⁸Between CFU/g feeds, suitably between about 7.5 × 10⁴CFU/g is raised Expect to about 1 × 10⁷Between CFU/g feeds.

In one embodiment, DFM is by more than about 1 × 10³CFU/g feeds are suitably more than about 1 × 10⁴CFU/g Feed is suitably more than about 7.5 × 10⁴CFU/g feeds are added in feed.

Expediently, the dosage of DFM can be between about 2 × 10 in feed addictive composition²CFU/g compositions to about 1 × 10¹⁰Between CFU/g compositions, suitably between about 1 × 10³CFU/g compositions are to about 1 × 10¹⁰Between CFU/g compositions, fit Preferably between about 1 × 10⁴CFU/g compositions are to about 1 × 10⁹Between CFU/g compositions, suitably between about 3.75x10⁴CFU/g Composition is to about 3x10⁸Between CFU/g compositions.

In one embodiment, DFM is by more than about 1 × 10⁵CFU/g compositions are suitably more than about 1 × 10⁶CFU/ G compositions are suitably more than about 3.75 × 10⁷CFU/g compositions are added in feed addictive composition.

The term as used herein " CFU " means Colony Forming Unit, and is the measurement of living cells, and wherein bacterium colony represents source From the cell aggregation of single progenitor cells.

Phytase

The term as used herein " phytase " refers to the phosphoric acid that catalysis includes phytate and phytic acid Ester hydrolysis and the enzyme (i.e. the polypeptide with phytase activity) for discharging inorganic phosphate.

For the present invention phytase can be divided into 6-phytase (being classified as E.C.3.1.3.26) or 3-Phytase (is classified as E.C.3.1.3.8).

In one embodiment, phytase can be 6-phytase (E.C.3.1.3.26).

In a preferred embodiment, the phytase for the present invention can be one in following commercial product One or more phytases in planting or being a variety of：

The term as used herein shares gene and means that the DNA vector for inverting biological body contains the conjunction based on consensus sequence At phytic acid enzyme gene, from the URA genes of non-pathogenic yeast saccharomyces cerevisiae and answering for escherichia coli plasmid pBR322 Starting point processed.

In one embodiment, phytase is Citrobacter phytase, is originated from：Such as Disclosed in WO2006/038062 (being hereby incorporated by reference) and WO2006/038128 (being hereby incorporated by reference) not Family name's citric acid bacterium (Citrobacter freundii), preferably Citrobacter freundii NCIMB 41247 and its variant；WO Bu Shi citric acid bacteriums YH-15 disclosed in 2004/085638；Bu Shi citric acids disclosed in WO2006/037328 are thin Bacterium ATCC51113 (being hereby incorporated by reference) and its in such as WO2007/112739 (being hereby incorporated by reference) and Variant disclosed in WO2011/117396 (being hereby incorporated by reference)；WO2006037327 (is hereby incorporated by reference) Disclosed in without malonic acid citric acid bacterium (Citrobacter amalonaticus), preferably without malonic acid citric acid bacterium ATCC 25405 or without malonic acid citric acid bacterium ATCC 25407；Institute is public in WO2006037327 (being hereby incorporated by reference) The Ji Shi citric acid bacteriums (Citrobacter gillenii) opened, preferably Ji Shi citric acid bacteriums DSM13694；Or intermediate lemon Lemon acetic bacterial (Citrobacter intermedius), kirschner citric acid bacterium (Citrobacter koseri), muroid lemon Acetic bacterial (Citrobacter murliniae), rodent citric acid bacterium (Citrobacter rodentium), Sai Lake Family name's citric acid bacterium (Citrobacter sedlakii), fertile gram Mans citric acid bacterium (Citrobacter werkmanii), The polypeptide or its variant of poplar Ge Shi citric acid bacteriums (Citrobacter youngae), citric acid bacterium species.

In one embodiment, phytase can come from Citrobacter, such as from Freund lemon The phytase of acetic bacterial, the phytase as taught in WO2006/038128, the bibliography are incorporated herein As reference.

In preferred embodiments, phytase is preferably by Danisco A/S BJ Rep Office with trade name Phyzyme XP^TMThe escherichia coli phytase of sale.

Alternatively, phytase can be Buttiauxella phytase, such as Buttiauxella agrestis (Buttiauxella agrestis) phytase, such as WO 2006/043178, WO2008/097619, WO2009/ 129489, it is taught in WO2006/038128, WO2008/092901, PCT/US2009/41011 or PCT/IB2010/051804 The phytase led, all patents are hereby incorporated by reference.

On the one hand, enzyme used is polypeptide shown in BP17 or SEQ ID No.1 or SEQ ID No.2 or its change Body, such as therewith at least 70% homogeneity, preferably therewith at least 75% homogeneity, preferably therewith at least 80% together One property, preferably therewith at least 85% homogeneity, preferably therewith at least 90% homogeneity, preferably therewith at least 95% homogeneity, preferably therewith at least 96% homogeneity, preferably therewith at least 97% homogeneity, preferably have therewith At least 98% homogeneity, the sequence preferably therewith at least 99% homogeneity.BP17 is six phosphorus of Buttiauxella species inositol The enzyme variants of sour enzyme, and be described in such as WO2008/097619, which is hereby incorporated by reference.

In one embodiment, enzyme used is BP17, and is described in such as WO2008/097619.BP17 is cloth The enzyme variants of mound Salmonella species phytase.

The sequence (excluding signal peptide) of BP17 is shown as SEQ ID No.1.The sequence for including signal peptide (underscore) of BP17 Row are shown as SEQ ID No.2.

SEQ ID NO：1：

NDTPASGYQVEKVVILSRHGVRAPTKMTQTMRDVTPNTWPEWPVKLGYITPRGEHLISLMGGFYRQKFQ QQGILSQGSCPTPNSIYVWTDVAQRTLKTGEAFLAGLAPQCGLTIHHQQNLEKADPLFHPVKAGICSMDKTQVQQAV EKEAQTPIDNLNQHYIPSLALMNTTLNFSKSPWCQKHSADKSCDLGLSMPSKLSIKDNGNEVSLDGAIGLSSTLAEI FLLEYAQGMPQAAWGNIHSEQEWALLLKLHNVYFDLMERTPYIARHKGTPLLQAISNALNPNATESKLPDISPDNKI LFIAGHDTNIANIAGMLNMRWTLPGQPDNTPPGGALVFERLADKSGKQYVSVSMVYQTLEQLRSQTPLSLNQPAGSV QLKIPGCNDQTAEGYCPLSTFTRVVSQSVEPGCQLQ

SEQ ID NO：2：

MTISAFNRKKLTLHPGLFVALSAIFSLGSTAYANDTPASGYQVEKVVILSRHGVRAPTKMTQTMRDVTP NTWPEWPVKLGYITPRGEHLISLMGGFYRQKFQQQGILSQGSCPTPNSIYVWTDVAQRTLKTGEAFLAGLAPQCGLT IHHQQNLEKADPLFHPVKAGICSMDKTQVQQAVEKEAQTPIDNLNQHYIPSLALMNTTLNFSKSPWCQKHSADKSCD LGLSMPSKLSIKDNGNEVSLDGAIGLSSTLAEIFLLEYAQGMPQAAWGNIHSEQEWALLLKLHNVYFDLMERTPYIA RHKGTPLLQAISNALNPNATESKLPDISPDNKILFIAGHDTNIANIAGMLNMRWTLPGQPDNTPPGGALVFERLADK SGKQYVSVSMVYQTLEQLRSQTPLSLNQPAGSVQLKIPGCNDQTAEGYCPLSTFTRVVSQSVEPGCQLQ

In one embodiment, phytase is AxtraPHY^TMPhytase.

AxtraPHY^TMPhytase is referred to herein as BP17 phytases.

In one embodiment, phytase can come from Hafnia, such as from honeycomb Hough Buddhist nun The phytase of bacterium (Hafnia alvei), the phytase as taught in US2008263688, this is with reference to text It offers and is hereby incorporated by reference.

In one embodiment, phytase can come from aspergillus, such as the inositol six from aspergillus oryzae Phosphatase.

In one embodiment, phytase can come from Penicillium, such as the inositol from penicillium funiculosum Six-phosphatase.

Unless the context clearly dictates otherwise, term " phytase " can be a kind of phytase or inositol six The combination of phosphatase.

Preferably, phytase is with about 1500FTU/kg to about 20000FTU/kg feeds, suitably about 1500FTU/ Kg to about 10000FTU/kg feeds, suitably about 1500FTU/kg feeds to about 5000FTU/kg feeds, suitably about 1500FTU/kg feeds are to about 3000FTU/kg feeds, more preferably from about 2000FTU/kg feeds to about 20000FTU/kg feeds, suitable Preferably about 2000FTU/kg feeds to about 10000FTU/kg feeds, suitably about 2000FTU/kg feeds to about 5000FTU/kg The range of feed, suitably about 2000FTU/kg feeds to about 3500FTU/kg feeds is present in feed.

In one embodiment, phytase is by more than about 1500FTU/kg feeds, be suitably more than about 2000FTU/kg feeds are suitably present in feed more than about 3000FTU/kg feeds.

In one embodiment, phytase by less than about 15000FTU/kg feeds, suitably less than about 12000FTU/kg feeds are present in feed.

In one embodiment, phytase is deposited with the range of about 1500FTU/kg to about 5000FTU/kg feeds It is in feed.

In one embodiment, phytase is deposited with the range of about 2000FTU/kg to about 5000FTU/kg feeds It is in feed.

In preferred embodiments, phytase is deposited with the range of about 1500FTU/kg to about 3000FTU/kg feeds It is in feed.

In a more preferred embodiment, phytase is with about 2000FTU/kg to the model of about 3000FTU/kg feeds It encloses and is present in feed.

In one embodiment, phytase can be by existing below in feed addictive composition：

A. the composition in adding to (such as dosage of manufacturer's recommended) feed is equal to or more than 50g/ tonnes of (MT) feeds When, by about 30,000FTU/g compositions or more；

B. the composition in adding to (such as dosage of manufacturer's recommended) feed is equal to or more than 75g/ tonnes of (MT) feeds When, by about 20,000FTU/g compositions or more；

C. the composition in adding to (such as dosage of manufacturer's recommended) feed is equal to or more than 100g/ tonnes of (MT) feeds When, by about 15,000FTU/g compositions or more；

D. the composition in adding to (such as dosage of manufacturer's recommended) feed is equal to or more than 100g/ tonnes of (MT) feeds When, by about 15,000FTU/g compositions or more；

E. the composition in adding to (such as dosage of manufacturer's recommended) feed is equal to or more than 150g/ tonnes of (MT) feeds When, by about 10,000FTU/g compositions or more；

F. the composition in adding to (such as dosage of manufacturer's recommended) feed is equal to or more than 200g/ tonnes of (MT) feeds When, by about 7,500FTU/g compositions or more；

G. the composition in adding to (such as dosage of manufacturer's recommended) feed is equal to or more than 300g/ tonnes of (MT) feeds When, by about 5,000FTU/g compositions or more.

Expediently, when phytase is present in dosage given above in feed addictive composition, directly Microorganism is fed with from 2.5x10³CFU DFM:1FTU enzymes are to 6.7x10⁶CFU:The range of 1FTU enzymes, suitably with from 3.8x10³CFU DFM:1FTU enzymes are to 2.0x10⁵CFU:The range of 1FTU enzymes is present in feed addictive composition.

It should be understood that as used herein, 1FTU (phytase unit) is defined as in 2009 phytases of ISO It measures under the reaction condition defined in (standard test for measuring phytase activity), from substrate in one minute The enzyme amount needed for 1 μm of ol inorganic orthophosphate is discharged, and 1FTU can be found in international standard ISO/DIS 30024:1-17,2009.

In one embodiment, expediently, being classified to enzyme using above-mentioned E.C. classification, and E.C. classification instructions It is teaching herein for measuring test in the measurement of 1FTU when there is the active enzyme.

Advantage

The interaction of DFM and enzyme is complicated, and is not intended to be limited to theory, very it is surprising that we It can be seen that improvement of the individual to the resistance of necrotic enteritis.Before making the present invention, not yet instructed for the specific purposes DFM with The specific combination of high phytase (for example, as taught herein).

In addition or alternatively, for animal shows (such as feed conversion rate), add more than by 1500FTU/kg feeds When entering phytase, the combination of DFM (such as antipathogen DFM) and phytase, which exists, to act synergistically.Independent When supplementing phytase, there is the raising (i.e. the reduction of FCR) of performance under 500 and 1500FTU/kg feeds, in inositol There are the reduction of performance (raisings of FCR) when six-phosphatase dosage is more than about 1500FTU/kg feeds.This is in necrotic enteritis It is especially true under conditions of attack.

But when combining DFM with phytase more than 1500FTU/kg feeds, and phytic acid is only used Enzymatic treatment is compared, and there are consistent numerical value to increase (i.e. the reduction of FCR) for performance.In 3000FTU/kg feeds and 10,000FTU/kg Between the level of feed, there are the reductions of big numerical value, lead to FCR minimum in all processing groups.Astoundingly, DFM is (especially Antipathogen DFM) with more than 1500FTU/kg feeds phytase combination generation synergistic combination.

It is one of the advantages of the invention that DFM (such as antipathogen DFM) and the combination of high dose phytase are aobvious Writing improves the effect of phytase improves animal performance and/or phosphorus reservation in individual.

Another advantage of the present invention is that DFM (such as antipathogen DFM) and the combination of high dose phytase change The phosphorus (such as diet phosphorus) being apt in individual absorbs and retains.This compound action is entirely unexpected.

It is one of the advantages of the invention that the feed addictive composition of the present invention can be to avoid the negative of necrotic enteritis The resistance for acting on or can be used for improving individual to necrotic enteritis.

Be not intended to be limited to theory, phytic acid enzymatic phytate (phosphorus in cereal and beans it is main Storage form) it is sequentially hydrolyzed into the lower myo-inositol derivative of phosphorylation degree, while discharging inorganic phosphate.Phytic acid The hydrolysis of salt, which causes to lose the endogenous of the amino acid of enteric cavity, to be reduced.Endogenous Amino Acids loss in intestines, which is reduced, reduces use In the availability of the nitrogen of bacterial growth, this contributes to the activity of DFM inhibition C.perfringens and other pathogenic bacteria.

It is not intended to be limited to theory, phytase is added and six phosphate of hydrolysis of inositol can lead to small intestine (12 Duodenum 12, jejenum and/or ileum) pH improve.Phytase can also actually increase species Escherichia coli and aerogenesis The amount of bacteria of capsular clostridium.This can be related to higher pH.Present invention has found that being added with high dose phytase DFM (especially antipathogen DFM) can offset the increase of pathogenic bacteria.

High dose phytase increases the hydrolysis of six phosphoric acid of animal intestinal tract mysoinositol, can to improve phosphorus, calcium, Energy, protein and other nutrients digestibility and animal may be showed advantageous.But astoundingly, the present inventor It has been found that there is clinical or subclinical enteropathy attack (such as the necrotic enteritis in broiler chicken, often in commercial animal production Occur) when, the phytase of high dose cannot reach the potential improvement to Nutrient digestibility and animal performance.Passing through will High dose phytase is combined with antipathogen DFM, surprisingly, it was found that even if in six phosphorus of very high inositol Sour enzyme level (such as 10,000FTU/kg feeds), animal performance also further enhance.

Combination and/or DFM with DFM and standard (low) phytic acid enzyme dosage are individually and/or enzyme (or even high dose flesh Alcohol six-phosphatase) individually to compare, the combination of high dose phytase and DFM, which astoundingly provide, reduces pathogen And/or the battalion in necrotic enteritis and/or feed conversion rate, and/or weight gain and/or individual performance and/or reduction muck Support significantly improving for object excretion.

The preparation of DFM and enzyme

DFM and one or more enzymes can be prepared in any suitable manner, with ensure preparation include living DFM and it is a kind of or Multiple active enzymes.

In one embodiment, DFM and one or more enzymes can be formulated as liquid, dry powder or particle.

Dry powder or particle can by means well known by persons skilled in the art, for example, in top-jet-type fluid bed coater, In Wurster type bottom spraying types fluid bed (buttom spray Wurster) or pass through rotary drum granulation (such as high shear system Grain), squeeze out, pan coating or prepared in micro- Component mixer.

For some embodiments, (such as being encapsulated) DFM and/or enzyme can be coated with.Expediently, DFM and enzyme can be formulated in In same coating or it is encapsulated in same capsule.Alternatively, one or more enzymes can be formulated in a coating or is encapsulated in In one capsule, and can be separately formulated in coating by DFM and one or more enzymes.In some embodiments, if such as DFM can generate endospore, then can provide DFM without any coating.In these cases, can simply by DFM endospores mix with one or more enzymes.In the latter case, (such as being encapsulated) one or more enzymes can be coated with.

In one embodiment, coating protects one or more enzymes from heat affecting and can be considered anti-thermit powder.

In one embodiment, feed addictive composition is configured to dry powder or particle, such as WO2007/044968 Described in (being known as TPT particles) or WO1997/016076 or WO1992/012645 (each patent is hereby incorporated by reference).

In one embodiment, feed addictive composition can be configured to the particle for fodder compound, wrapped It includes：Core；Activating agent；With at least one coating, after one or more conditions in following, the activating agent of particle retains At least 50% activity, at least 60% activity, at least 70% activity, at least 80% activity：A) feed pill method, b) steam heating Feed preprocess method, c) storage, d) as in the mixture without pill ingredient storage and e) conduct include at least one The feed base mix of kind compound chosen from the followings or the ingredient storage in feed premix：Trace minerals, organic acid, The compound of reduced sugar, vitamin, choline chloride and generation acidity or alkaline feed basic mixture or feed premix.

About particle, at least one coating may include at least moisture hydrating material of 55w/w% for constituting particle；And/or At least one coating may include two coatings.Two coatings can be moisture hydration coating and moisture barrier coating.At some In embodiment, moisture hydration coating can be the 25w/w% of particle between 60w/w% and moisture barrier coating can be particle 2w/w% between 15w/w%.Moisture hydration coating can be selected from inorganic salts, sucrose, starch and maltodextrin and moisture screen Barrier coating can be selected from polymer, natural gum, whey and starch.

Particle can be used feed pill method prepare and feed preprocess method can be carried out between 70 DEG C to 95 DEG C to Between most minutes, such as 85 DEG C to 95 DEG C.

In one embodiment, feed addictive composition can be configured to the particle for animal feed comprising： Core；Activating agent, the activating agent of particle after storage and particle be ingredient steam heat pill method after retain to Few 80% activity；Moisture barrier coating；Moisture is hydrated coating, is at least 25w/w% of particle, and pill method is heated in steam Before, which has the water activity less than 0.5.

It can be inorganic salts that particle, which can have the moisture barrier coating selected from polymer and natural gum and moisture hydrating material,.Water Point hydration coating can be particle 25w/w% between 45w/w% and moisture barrier coating can be the 2w/w% of particle extremely Between 10w/w%.

Steam heating pill method can be used to prepare for particle, and this method can be carried out within several points between 85 DEG C to 95 DEG C Clock.

In some embodiments, diluent (such as starch powder, lime stone or the like) can be used to dilute DFM (examples Such as DFM endospores).

In one embodiment, composition is in edible liquid preparation, it is preferable that this liquid edible object Containing one or more in following：Buffer, salt, sorbierite and/or glycerine.

In another embodiment, can by by one or more enzymes apply (such as spraying) in carrier substrate (such as The wheat pulverized) come up formula feed compositions of additives.

In one embodiment, feed addictive composition of the invention can be formulated as pre-composition.It is only used as example, it should Pre-composition may include one or more feed ingredients, such as one or more minerals and/or one or more vitamins.

In one embodiment, it can will be selected from such as at least one for the DFM of the present invention and/or one or more enzymes At least one of lower physiologically acceptable carrier is prepared together：Maltodextrin, lime stone (calcium carbonate), ring paste Essence, wheat or wheat component, sucrose, starch, Na₂SO₄, talcum powder, PVA, sorbierite, benzoate, sorbate, glycerine, sugarcane Sugar, propylene glycol, 1,3- propylene glycol, glucose, p-hydroxybenzoate, sodium chloride, citrate, acetate, phosphate, calcium, Metabisulfite, formates and its mixture.

Packaging

In one embodiment, the feed addictive composition to the present invention and/or pre-composition and/or feed wrap Dress.

In a preferred embodiment, by feed addictive composition and/or pre-composition and/or feed package in bag In (such as paper bag).

In alternative embodiment, feed addictive composition and/or pre-composition and/or feed can be sealed in container In.The container of any suitable can be used.

Feed

The feed addictive composition of the present invention can be used as feed or be used to prepare feed.

Term " feed (feed) " is used with " feed (feedstuff) " herein as synonym.

Depending on purposes and/or application model and/or method of application, feed can be in solution form or be in solid form.

When as (or being used to prepare) feed (such as functional feed), the present composition can in following substance One or more combined uses：Nutritionally acceptable carriers, nutritionally acceptable diluent, nutritionally acceptable figuration Agent, nutritionally acceptable adjuvant, neutraceutical active ingredients.

In preferred embodiments, the feed addictive composition of the present invention is mixed with feed ingredient to form feed.

The term as used herein " feed ingredient " means all or part of feed.The part of feed can refer to the one of feed More than one (such as 2 kinds or 3 kinds or 4 kinds) ingredients of kind ingredient or feed.In one embodiment, term " feed ingredient " Cover pre-composition or premix ingredients.

Preferably, feed can be forage (fodder) or its pre-composition, mixed feed or its pre-composition.In an embodiment party In case, can the feed addictive composition of the present invention be mixed or be mixed to compound with mixed feed, mixed feed component The pre-composition of feed is mixed into the pre-composition of forage, forage component or forage.

The term as used herein forage means the arbitrary food for being supplied to animal (and non-animal oneself must look for food).Forage Cover cut plant.

Term forage includes feed, oil and mixed ration and germinated ceral and the beans of ensilage, compacting and pill.

Forage can be obtained from one or more in plant chosen from the followings：Barley, rape (Canola), corn (beautiful another name for Sichuan Province Broomcorn millet), grain, oat, sorghum, soybean, wheat and beans.

Term " mixed feed " means the commercial feed of coarse powder, pellet, nut, cake or debris form.Mixed feed can be by Plurality of raw materials and additive are blended.These blends are prepared according to the specific requirements of target animal.

Mixed feed can be to provide the complete feeding-stuffs of all daily needed nutrients, provide daily ration (ration) (albumen Matter, energy) a part concentrate or the replenishers of additional micronutrient (such as minerals and vitamins) are only provided.

It is feed grain for the main component in mixed feed comprising corn and soybean, sorghum, wheat, oat and big Wheat.

Expediently, the composition that the pre-composition mentioned by this paper can be made of micro constitutent, which is example As vitamin, minerals, chemical preservative, antibiotic, tunning and other must ingredient.Pre-composition is generally suitable in altogether The composition being mixed into business daily ration.

The arbitrary feed of the present invention may include one or more Feed Materials chosen from the followings：A) cereal, such as granule paddy Object (such as wheat, barley, rye, oat and combinations thereof) and/or big grain cereal such as maize or sorghum；B) pair from cereal Product, such as corn protein powder, wet cake (wet cake for being based especially on corn), distiller's dried grain (DDG) (are based especially on the dry of corn Vinasse (cDDG)), distiller's dried grain soluble matter (DDGS) (the distiller's dried grain soluble matter (cDDGS) for being based especially on corn), wheat bran, wheat Coarse powder, wheat wheat-middlings, rice bran, rice husk, oat shell, palm kernel and citrus pulp；C) it is obtained from the protein in following source：As soybean, Sunflower, peanut, lupin, pea, broad bean, cotton, Canola, fish meal, dry plasma protein, meat and bone meal, potato egg In vain, whey, copra, sesame；D) it is obtained from the oil ＆ fat in plant and animal source；E) minerals and vitamins.

The feed of the present invention contains the corn of at least 10wt%, at least 20wt%, at least 30wt% or at least 50wt% With soy meal or corn and full-fat bean or wheat flour or sunflower powder.

The feed of the present invention can include the corn DDGS between about 0% to about 40%.If feed includes arbitrary corn DDGS, then it can include the corn DDGS between about 5% to about 40%.For poultry, when there are corn DDGS, feed is flat It can include the corn DDGS between about 7% to about 15%.For pig, when there are corn DDGS, feed can averagely wrap Containing 5% to 40% corn DDGS.

The feed of the present invention can be comprising corn as single cereal, in this case, and feed can include about 35% To about 80% corn.

In the present invention, one or more during feed can be as follows：Mixed feed and pre-composition comprising pellet, Nut or (domestic animal use) cake；Crops or crop residues：Corn and soybean, sorghum, oat, barley, copra, husk, Sugar beet waste products；Fish meal；Meat and bone meal；Molasses；Oil cake and filter press cake；Oligosaccharide；The forage plant of preservation：Ensilage；Sea Algae；Seed and cereal are entire or by the preparations such as rolling or grinding；Germinated ceral and beans；Yeast extract.

In some embodiments, the term feed in the present invention is also covered by pet food.Pet food is intended to by doting on The edible plant of object or animal material, such as dog food or cat foods.Pet food (such as dog and cat foods) can be drying Form (grate foods as being used for dog) or wet canned form.Cat foods can contain amino acid taurine.

In some embodiments, the term feed in the present invention is also covered by fish food.Fish food, which usually contains, makes cultivation Fish is kept fit required main nutrient, trace element and vitamin.Fish food can be the form of small pieces, pellet or piece. The form (some of which rapid subsidence) for being pressed into pellet is frequently used for larger fish or species are raised at bottom.Some fish foods also contain The additive of such as bata-carotene or sex hormone etc, manually to enhance the color of ornamental value fish.

In some embodiments, the term feed in the present invention is also covered by birdseed product.Birdseed product include being used for bird feeder In and the food for feeding pet bird.In general, birdseed product are made of a variety of seeds, but it can also cover leaf fat (ox or suet Fat).

The term as used herein " contact " refers to that the composition of the present invention is directly or indirectly applied to product (such as to raise Material) in.The example of workable applying method includes but not limited to：Production is handled in the material comprising feed addictive composition Product, by the way that feed addictive composition and product mix are directly applied, feed addictive composition is sprayed into product table It is immersed in the preparation of feed addictive composition on face or by product.

In one embodiment, it is preferred to which the feed addictive composition of the present invention is mixed with product (such as feed). Alternatively, it may include feed addictive composition in the lotion or primitive component of feed.

For some application, it is important that make composition on the product surface for waiting for influence/processing can with or can be used for waiting for The product surface of influence/processing.It is one or more in following advantageous feature that this enables composition to assign：Show benefit.

Can apply the present invention feed addictive composition so that product (such as primitive component of feed or feed) spread, It is coated with and/or is impregnated with the DFM of controlled quentity controlled variable and one or more enzymes.

DFM and one or more enzymes can be used simultaneously (for example, when they are mixed or even passing through not at them When being delivered with approach) or sequentially use (for example, they can be delivered by different approaches).In one embodiment, it is preferred to ground, Apply DFM and one or more enzymes simultaneously.Preferably, DFM is mixed with one or more enzymes, they is then delivered to feed Or the primitive component of feed.

The present invention feed addictive composition in DFM can be suitable concentration addition --- such as final feed product In concentration can provide about 2 × 10⁵CFU to about 2 × 10¹¹Between CFU, suitably between about 2 × 10⁶CFU to about 1 × 10¹⁰CFU Between, suitably between about 3.75 × 10⁷CFU to about 1 × 10¹⁰Daily dose between CFU.

Preferably, feed addictive composition of the invention will be to a maximum of about of 70 DEG C, a maximum of about of 85 DEG C or a maximum of about of 95 DEG C Heat treatment be heat-staple.Heat treatment can carry out most about 1 minute, most about 5 minutes, most about 10 minutes, most about 30 minutes, most about 60 minutes.Term is heat-staple to be meant to exist in additive before being heated to assigned temperature/and it is active In enzyme component and/or DFM at least about 75% after cooling to room temperature there are still/it is active.Preferably, it is being heated to specified temperature Exist in additive before degree and active enzyme component and/or DFM at least about 80% after cooling to room temperature there are still And it is active.

In especially preferred embodiment, feed addictive composition is homogenized to generate powder.

In alternative preferred embodiment, feed addictive composition is configured to as described in WO2007/044968 Particle (be referred to as TPT particles), which is hereby incorporated by reference.

In another preferred embodiment of the present, when feed addictive composition is configured to particle, these particles include to apply Hydrated barrier salt of the cloth on protein core.The advantage of this salt coating is to improve heat resistance, improve storage stability and guarantor Protect the influence of other feed addictives to enzyme and/or DFM from script with detrimental effect.

Preferably, the salt for being used for salt coating has the water activity or constant wet more than 60% for being more than 0.25 at 20 DEG C Degree.

Preferably, salt coating includes Na₂SO₄。

The method for preparing feed addictive composition may also include another step of powder pill.Powder can be with ability Other components known to domain are mixed.Powder or mixture comprising powder can be forced by mold, and by obtained material Item is cut into the suitable pellet of variable-length.

Optionally, pill step may include the steam treatment carried out before forming pellet or conditioning stage.Can will include The mixture of powder is placed in regulation machine, such as the mixing machine that carrying vapour sprays.Mixture is heated to specifying in regulation machine Temperature, such as from 60 DEG C to 100 DEG C, typical temperature can be 70 DEG C, 80 DEG C, 85 DEG C, 90 DEG C or 95 DEG C.Residence time is indefinite, can From the several seconds to several minutes or even a few hours.Such as 5 seconds, 10 seconds, 15 seconds, 30 seconds, 1 minute, 2 minutes, 5 minutes, 10 minutes, 15 points Clock, 30 minutes and 1 hour.

It should be appreciated that the feed addictive composition of the present invention is suitable for being added in any suitable Feed Material.

The term as used herein Feed Material refers to the basal feed material waited for by animal edible.It should also be understood that this may include For example, at least one or more crude cereal and/or processed plant and/or animal material, such as soy meal or bone Powder.

The term as used herein " feed " refers to being added to the feed material of one or more feed addictive compositions Material.

Technical staff is, it will be recognized that different animals needs different feeds or even same animal that can also need not Same feed, this depends on the purpose for raising the animal.

Preferably, feed may include such Feed Material, it includes maize or corn, wheat, barley, triticale, Rye, rice, cassava, sorghum and/or arbitrary by-product, and the ingredient rich in protein, such as soy meal, ground oil seeds, Kano Draw powder, cotton seed meal, sunflower seed powder, animal byproducts powder and its mixture.It is highly preferred that feed may include animal tallow and/or Vegetable oil.

Optionally, feed can also contain additional minerals (such as calcium) and/or additional vitamin.

Preferably, feed is corn soybean powder mixture.

In one embodiment, it is preferred to ground, feed is not pet food.

On the other hand, the method for producing feed is provided.Feed usually produces in feed metal, first in grinding machine Raw material is first ground into suitable granularity, is then mixed with additive appropriate.Feed can then be used as thickener or pellet system It is standby；The latter, which is usually directed to, to be raised the temperature to target level and feed is then made to pass through mold with the pellet that generates specific dimensions Method.Pellet is set to cool down.It then, can adding liquid additive such as fat and enzyme.The preparation of feed may also refer to additional step, Its extrusion or expansion before being included in pill --- it is carried out especially by the appropriate technology that may include at least using steam.

Feed can be for nonruminant such as poultry (such as broiler chicken, laying hen, Breeder hens, turkey, duck, goose, aquatic bird), pig The feed of (all age categories), pet (such as dog, cat) or fish, preferably feed are used for poultry.

In one embodiment, feed is not intended to laying hen.

It is only used as example, the % that the feed for chicken (such as broiler chicken) can provide in such as according to the form below includes listed in following table It is one or more in ingredient：

Ingredient	Opened guideway (%)	Fattening material (%)
			Maize	46.2	46.7
Sizing	6.7	10.0
			Maize DDGS	7.0	7.0
Soy meal 48%CP	32.8	26.2
			Animal/vegetable fat blend	3.0	5.8
L-lysine HCl	0.3	0.3
			DL- methionines	0.3	0.3
L-threonine	0.1	0.1
			Salt	0.3	0.4
Lime stone	1.1	1.1
			Dicalcium Phosphate	1.2	1.2
Poultry vitamin and trace mineral	0.3	0.3

It is only used as example, the diet specification for chicken (such as broiler chicken) can be as shown in following table：

Diet specification
			Crude protein (%)	23.00	20.40
Poultry metabolic energy (kcal/kg)	2950	3100
			Calcium (%)	0.85	0.85
Available phosphorus (%)	0.38	0.38
			Sodium (%)	0.18	0.19
Digestible lysine (%)	1.21	1.07
			Digestible methionine (%)	0.62	0.57
Digestible methionine+cysteine (%)	0.86	0.78
			Digestible threonine (%)	0.76	0.68

It is only used as example, the feed for laying hen can be made of one or more in ingredients listed in following table, such as with The % provided in following table：

Ingredient	Laying period (%)
		Maize	10.0
Wheat	53.6
		Maize DDGS	5.0
Soy meal 48%CP	14.9
		Sizing	3.0
Soybean oil	1.8
		L-lysine HCl	0.2
DL- methionines	0.2
		L-threonine	0.1
Salt	0.3
		Dicalcium Phosphate	1.6
Lime stone	8.9
		Poultry vitamin and trace mineral	0.6

It is only used as example, the diet specification for laying hen can be as shown in following table：

Diet specification
		Crude protein (%)	16.10
Poultry metabolic energy (kcal/kg)	2700
		Lysine (%)	0.85
Methionine (%)	0.42
		Methionine+cysteine (%)	0.71
Threonine (%)	0.60
		Calcium (%)	3.85
Available phosphorus (%)	0.42
		Sodium (%)	0.16

It is only used as example, the feed for turkey can be made of one or more in ingredients listed in following table, such as with The % provided in following table：

It is only used as example, the diet specification for turkey can be as shown in following table：

Diet specification
					Crude protein (%)	29.35	26.37	22.93	20.00
Poultry metabolic energy (kcal/kg)	2.850	2.900	2.950	3.001
					Calcium (%)	1.43	1.33	1.22	1.02
Available phosphorus (%)	0.80	0.71	0.65	0.53
					Sodium (%)	0.16	0.17	0.17	0.17
Digestible lysine (%)	1.77	1.53	1.27	1.04
					Digestible methionine (%)	0.79	0.71	0.62	0.48
Digestible methionine+cysteine (%)	1.12	1.02	0.90	0.74
					Digestible threonine (%)	1.03	0.89	0.73	0.59

It is only used as example, the feed for piglet can be made of one or more in ingredients listed in following table, such as with The % provided in following table：

Ingredient	Stage 1 (%)	Stage 2 (%)
			Maize	20.0	7.0
Wheat	25.9	46.6
			Rye	4.0	10.0
Sizing	4.0	4.0
			Maize DDGS	6.0	8.0
Soy meal 48%CP	25.7	19.9
			Dry whey	10.0	0.0
Soybean oil	1.0	0.7
			L-lysine HCl	0.4	0.5
DL- methionines	0.2	0.2
			L-threonine	0.1	0.2
L-Trp	0.03	0.04
			Lime stone	0.6	0.7
Dicalcium Phosphate	1.6	1.6
			Pig vitamin and trace mineral	0.2	0.2
Salt	0.2	0.4

It is only used as example, the diet specification for piglet can be as shown in following table：

Diet specification
			Crude protein (%)	21.50	20.00
Pig digestible energy (kcal/kg)	3380	3320
			Pig net energy (kcal/kg)	2270	2230
Calcium (%)	0.80	0.75
			Digestible phosphorus (%)	0.40	0.35
Sodium (%)	0.20	0.20
			Digestible lysine (%)	1.23	1.14
Digestible methionine (%)	0.49	0.44
			Digestible methionine+cysteine (%)	0.74	0.68
Digestible threonine (%)	0.80	0.74

It is only used as example, the feed for grower pigs/fattening pig can be by one or more groups in ingredients listed in following table At, such as the % that is provided in following table：

Ingredient	Growth material/fattening material (%)
		Maize	27.5
Soy meal 48%CP	15.4
		Maize DDGS	20.0
Wheat bran	11.1
		Rice bran	12.0
Canola seed powder	10.0
		Lime stone	1.6
Dicalcium Phosphate	0.01
		Salt	0.4
Pig vitamin and trace mineral	0.3
		Lysine-HCl	0.2
Vegetable oil	0.5

It is only used as example, the diet specification for grower pigs/fattening pig can be as shown in following table：

Diet specification
		Crude protein (%)	22.60
Pig metabolic energy (kcal/kg)	3030
		Calcium (%)	0.75
Available phosphorus (%)	0.29
		Digestible lysine (%)	1.01
Digestible methionine+cysteine (%)	0.73
		Digestible threonine (%)	0.66

Form

The feed addictive composition and other components and/or can be with any suitable shape comprising their feed of the present invention Formula uses.

The feed addictive composition of the present invention can be used with solid or liquid preparation or its alterative version.Solid pharmaceutical preparation Example includes powder, paste, bolus, capsule, pill, tablet, powders and granules, can be wettable, spraying It is dry or freeze-drying.The example of liquid preparation is including but not limited to aqueous, organic or aqueous-organic solutions agent, is suspended Agent and emulsion.

In some applications, can the DFM of the present invention or feed addictive composition be mixed or is applied to feed and drunk In water.In one embodiment, the dosage range including in water inlet is about 1 × 10³CFU/ animals/day is to about 1 × 10¹⁰CFU/ Animal/day, more preferably from about 1 × 10⁷CFU/ animals/day.

The example of suitable form include it is following in it is one or more：Powder, paste, bolus, pellet, tablet, ball Agent, capsule, vaginal suppository, solution or suspension can contain corrigent or colorant, for discharging at once, postponing to release It puts, adjust release, sustained release, pulse release or controlled release application.

For example, if the composition of the present invention is used with solid (such as pellet form), can also contain as follows In it is one or more：Excipient, such as microcrystalline cellulose, lactose, sodium citrate, calcium carbonate, calcium monohydrogen phosphate and glycine；It collapses Solve agent, such as starch (preferably corn, potato or tapioca), sodium starch glycollate, croscarmellose sodium and Certain composition silicates；Granulation binders, such as polyvinylpyrrolidone, hydroxypropyl methyl cellulose (HPMC), hydroxypropyl are fine Dimension plain (HPC), sucrose, gelatin and Arabic gum；Lubricant, such as magnesium stearate, stearic acid, Gan You behenic acids esters and talcum Powder.

The example for being used to prepare the nutritionally acceptable carriers of these forms includes (for example) water, salting liquid, alcohol, organic Silicon (silicone), wax, mineral jelly, vegetable oil, polyethylene glycol, propylene glycol, liposome, sugar, gelatin, lactose, amylose, Magnesium stearate, talcum powder, surfactant, silicic acid, viscous paraffin, perfume oil, fatty mono glyceride and fatty acid glycerine two Ester, petroleum ether (petroethral) aliphatic ester, methylol-cellulose, polyvinylpyrrolidone etc..

Preferred excipient for these forms includes lactose (lactose), starch, cellulose, lactose (milk ) or high molecular weight polyethylene glycol sugar.

For aqueous suspension and/or elixir, can by composition and various sweeteners or the corrigent of the present invention, Color substance or dyestuff, with emulsifier and/or suspending agent and with diluent (such as water, propylene glycol and glycerine) and their group Conjunction is combined.

Unwetted whey is commonly used as the carrier for DFM (especially bacterium DFM) and is the good medium for causing growth.

Paste containing bacterium DFM can be congealed into vegetable oil and inertia glue together with assignment system.

Fungal product can be formulated together with the cereal byproduct as carrier.

In one embodiment, it is preferred to ground, feed addictive composition according to the present invention is not microparticulate systems (example Microparticulate systems as taught in WO2005/123034) form.

It gives

The DFM and/or feed addictive composition of the present invention may be configured to once give or designed in terms of daily Feeding.

The optimised quantity for being ready to use in the composition (and each component therein) in present invention combination will be depending on pending Product and/or the method and/or its desired use for so that product is contacted with composition.

Amount for DFM and one or more enzymes in composition, which should be, to be enough effective for and is enough to keep to have enough Effectiveness is in the amount of the performance for the animal for improving feed product of the feeding containing the composition.

The time span of validity should extend up to at least with the product (such as feed addictive composition or contain Its feed) time.

In the range of the ratio of DFM and enzyme can be provided below in feed：

DFM：Phytase (CFU/FTU)：From 2.5x10³CFU DFM:1FTU enzymes are to 6.7x10⁶CFU:1FTU enzymes In the range of；It is preferred that from 3.8x10³CFU DFM:1FTU enzymes are to 2.0x10⁵CFU:In the range of 1FTU enzymes.

In one embodiment, it is preferred to ground, feed includes following：

(at least) phytase of 1500FTU/kg feeds；With

At least 7.5x10⁴CFU/g to 3.0x10⁵The Envivo Pro (DFM) of CFU/g feeds.

In one embodiment, it is preferred to ground, feed includes following：

(at least) phytase of 2000FTU/kg feeds；With

At least 7.5x10⁴CFU/g to 3.0x10⁵The Envivo Pro (DFM) of CFU/g feeds.

In one embodiment, it is preferred to ground, feed includes following：

(at least) phytase of 3000FTU/kg feeds；With

At least 7.5x10⁴CFU/g to 3.0x10⁵The Envivo Pro (DFM) of CFU/g feeds.

In one embodiment, it is preferred to ground, feed includes following：

(at least) phytase of 1500FTU/kg feeds；With

At least 5x10⁴CFU/g to 1x10⁷The DFM of CFU/g feeds.

In one embodiment, it is preferred to ground, feed includes following：

(at least) phytase of 2000FTU/kg feeds；With

At least 5x10⁴CFU/g to 1x10⁷The DFM of CFU/g feeds.

In one embodiment, it is preferred to ground, feed includes following：

(at least) phytase of 3000FTU/kg feeds；With

At least 5x10⁴CFU/g to 1x10⁷The DFM of CFU/g feeds.

In one embodiment, feed includes following：

In preferred embodiments, feed addictive composition is as follows feed is added comprising enough enzyme and DFM：

(at least) phytase of 1500FTU/kg (such as (at least) 2000FTU/kg) feed；With

75,000CFU/g to 150,000CFU/g feeds Envivo Pro (DFM).

(at least) phytase of 1500FTU/kg (such as (at least) 2000FTU/kg) feed；With

37,500CFU/g to 75,000CFU/g feeds Envivo Pro (DFM).

With the combination of other compositions

For in the present invention DFM and one or more enzymes can be applied in combination with other compositions.Therefore, the invention further relates to Combination.The combination of DFM and phytase can be described as " feed addictive composition of the invention " herein.

The combination of the present invention is comprising the feed addictive composition (or one or more component) of the present invention and suitable for dynamic Object is edible and another ingredient of medicine or physiological benefit can be provided for eater.

In one embodiment, it is preferred to ground, it is not other enzymes or other DFM to be somebody's turn to do " another seam part ".

These ingredients can be probiotics.Probiotics are typically the non-digestion degraded or absorbed not in upper digestive tract Property carbohydrate (oligosaccharide or polysaccharide) or sugar alcohol.It is neutralized for commercial product and includes for the known probiotics of the present invention Inulin (oligofructose or FOS) and oligomeric trans-galacto (GOS or TOS).Suitable probiotics include palatinose-oligosaccharides, It is soyabean oligosaccharides, alginate, xanthans, pectin, locust bean gum (LBG), inulin, guar gum, galactooligosaccharide (GOS), low Fructooligosaccharides (FOS), nondegradable starch, lactosucrose, lactulose, lactitol, maltitol, maltodextrin, dextrosan (i.e.), lactitol, lactosucrose, soyabean oligosaccharides, palatinose, oligoisomaltose, glucooligosaccharides sugar and it is oligomeric Xylose, pectin fragments, dietary fiber, Oligomeric manna sugar.

Dietary fiber may include non-starch polysaccharide (such as arabinoxylan), cellulose and many other plants Ingredient, such as resistant dextrin, inulin, lignin, wax, chitin, pectin, beta glucan and oligosaccharide.

In one embodiment, the present invention relates to the feed addictive composition of the present invention (or its one or more groups Point) with the combination of probiotics.In another embodiment, the present invention relates to include DFM and high dose phytase and benefit The feed addictive composition of the combination (or consisting essentially of or be made from it) of bacterium member.

Probiotics can be with feed addictive composition (or its component) of the invention simultaneously (for example, mixing therewith Object is delivered simultaneously by identical or different approach) or (such as by identical or different approach) is applied after which.

The present invention combination other compositions include dextrosan (such as) and/or maltodextrin and/or breast Sugar alcohol.These other compositions can be optionally added in feed addictive composition to assist drying process and help DFM Survival.

Other examples of other suitable ingredients include it is following in it is one or more：Thickener, gelling agent, emulsifier, Adhesive, crystal modifier, sweetener (including artificial sweetener), rheology modifier, stabilizer, antioxidant, dyestuff, enzyme, Carrier, medium, excipient, diluent, lubricant, corrigent, coloring material, suspending agent, disintegrant, granulation binders etc.. These other compositions can be natural.These other ingredients can be prepared by using chemistry and/or zymetology technology.

In a preferred embodiment, the DFM and/or enzyme being used in the present invention can be combined with one or more lipids It uses.

For example, the DFM and/or enzyme that are used in the present invention can be applied in combination with one or more lipid micelles.Lipid Protomere can be simple lipid protomere or complex lipids protomere.

Lipid micelles can be the aggregation of the orientation molecule of amphiprotic substance (such as lipid and/or oil).

The term as used herein " thickener or gelling agent " refer to by slow down or prevent particle (immiscible liquid it is small Drop, air or insoluble solid) it moves and prevents the product detached.Viscosity is caused to increase in individual hydrate molecule, to subtract When slow separation, thicken.When the molecule connection of hydration formed can trapped particle three-dimensional network, thus make these particles solid Timing is gelled.

The term as used herein " stabilizer " is defined as keeping product (such as feed product) from over time becoming The combination of the ingredient or ingredient of change.

The term as used herein " emulsifier " refers to the ingredient (such as feed ingredient) for preventing lotion from detaching.Lotion is two kinds Unmixing substance, one kind exist with droplet, are contained in another one.Lotion can (wherein droplet or dispersed phase be by oil-in-water Oil and continuous phase is water) or Water-In-Oil (wherein water becomes dispersed phase and continuous phase is oil) composition.It also can be by using emulsifier To make foam (it is gas-in-liquid) and suspension (it consolidates suspension for liquid) stablize.

The term as used herein " adhesive " refers to by physically or chemically reacting the combined ingredient (example of product Such as feed ingredient).For example, during " gelling ", water is absorbed, to provide in conjunction with effect.However, adhesive can absorb other Liquid (such as oil), to hold it in product.In the background of the invention, adhesive will be commonly used in solid or low water Point product (such as baked product：Dessert, baked donut, bread etc.) in.

" carrier " or " medium " means suitable for the material using DFM and/or enzyme, and includes known in the art It anticipates this material, such as arbitrary liquid, gel, solvent, liquid diluent, stabilizer etc., it is nontoxic and not with harmful way It interacts with any ingredient of composition.

The present invention is provided to prepare the method for feed addictive composition, this method includes by DFM and phytic acid Enzyme is mixed at least one selected from least one of following physiologically acceptable carrier：Maltodextrin, lime stone (calcium carbonate), cyclodextrin, wheat or Wheat components, sucrose, starch, Na₂SO₄, talcum powder, PVA, sorbierite, benzoate, mountain Pears hydrochlorate, glycerine, sucrose, propylene glycol, 1,3- propylene glycol, glucose, p-hydroxybenzoate, sodium chloride, citrate, acetic acid Salt, phosphate, calcium, metabisulfite, formates and its mixture.

The example of excipient include it is following in it is one or more：Microcrystalline cellulose and other celluloses, lactose, citric acid Sodium, calcium carbonate, calcium monohydrogen phosphate, glycine, starch, lactose and high molecular weight polyethylene glycol.

The example of disintegrant include it is following in it is one or more：Starch (preferably corn, potato or tapioca), hydroxyl Guanidine-acetic acid sodium starch, croscarmellose sodium and certain composition silicates.

The example of granulation binders include it is following in it is one or more：Polyvinylpyrrolidone, hydroxypropyl methyl fiber Plain (HPMC), hydroxypropyl cellulose (HPC), sucrose, maltose, gelatin and Arabic gum.

The example of lubricant include it is following in it is one or more：Magnesium stearate, stearic acid, Compritol 888 ATO and talcum Powder.

The example of diluent include it is following in it is one or more：Water, ethyl alcohol, propylene glycol and glycerine and combinations thereof.

Other compositions can be used simultaneously (such as when they are mixed or even when they pass through different approaches When delivering) or sequentially use (such as they can be delivered by different approaches).

Preferably, when mixing the feed addictive composition of the present invention with another ingredient, DFM remains living.

In one embodiment, it is preferred to ground, feed addictive composition of the invention does not include chromium or Organic Chromium.

Concentrate

Can be concentration form for the DFM in the present invention.In general, these concentrates include the DFM of suitable high concentration.

The content (colony forming unit, CFU) for the living cells that the feed addictive composition of the present invention can have is at least 10⁴CFU/g (includes suitably at least 10⁵CFU/g, such as at least 10⁶CFU/g, for example, at least 10⁷CFU/g, at least 10⁸CFU/g) extremely About 10¹⁰CFU/g (or even about 10¹¹CFU/g or about 10¹²CFU/g in the range of).

When DFM is concentration form, the content for the living cells that feed addictive composition of the invention can have is extremely Few 10⁹CFU/g to about 10¹²CFU/g, preferably at least 10¹⁰CFU/g to about 10¹²In the range of CFU/g.

Powder, granule and the liquid composition dilutable water of concentration form are resuspended in water or other are suitable dilute It releases in agent (such as growth medium appropriate, such as breast or minerals or vegetable oil), to generate available composition.

Can be prepared according to methods known in the art concentration form the present invention DFM or feed addictive composition or The combination of the present invention.

In one aspect of the invention, composition catalase or feed in a concentrated form.

The composition of the present invention can be spray-dried or is freeze-dried by methods known in the art.

Prepared with spray drying process particle typical method be related to being dissolved in appropriate solvent solid material (for example, DFM cultures in fermentation medium).Alternatively, material can be suspended or is emulsifiable in non-solvent to form suspension or breast Liquid.Can optionally add at this stage other compositions (as discussed above) or such as antimicrobial, stabilizer, dyestuff and Assist the ingredient of substance of drying process etc.

Then make solution atomization to form the mist of droplet.Droplet immediately enters in hothouse, wherein its contact drying Gas.Solvent evaporates into dry gas from droplet and cures droplet, to form particle.Then grain is detached from dry gas Son is simultaneously collected.

Individual

The term as used herein " individual " mean it is to be administered or applied the present invention feed addictive composition or comprising The animal of the feed of the feed addictive composition of the present invention.

The term as used herein " individual " means animal.

Preferably, individual is mammal, bird, fish or shellfish, including such as domestic animal or performing animal (such as dote on Object).

In one embodiment, " individual " is domestic animal.

The term as used herein " domestic animal " refers to arbitrary farm-animals.Preferably, domestic animal be it is following in it is one or more： Ruminant, such as ox (such as milk cow or bull (including calf))；Nonruminant, such as poultry (including broiler chicken, chicken and turkey), pig (including piglet), bird；Aquatic animal, such as fish, without stomach fish, have stomach fish, fresh-water fishes (such as salmon, gadus, trout and carp, such as Fancy carp), ocean fish (such as jewfish), shellfish (such as shrimp, mussel and scallop), horse (including horse racing), sheep (including lamb).

In another embodiment, " individual " is performing animal or pet or the animal that is maintained in the environment of zoo.

The term as used herein " performing animal or pet or the animal being maintained in the environment of zoo " refers to arbitrary correlation Animal, including canid (such as dog), felid (such as cat), rodent (such as cavy, rat, mouse), bird, Fish (including fresh-water fishes and ocean fish) and horse.

In one embodiment, individual can be attacked by enteropathogen.

As example, may exist one or more enteropathogens in individual intestines or alimentary canal.For example, individual intestines or The one or more enteropathogens that can have by such level in alimentary canal, the level：

I) lead to the loss of the performance of animal；And/or

Ii) it is clinical relevant levels；Or

Iii) it is subclinical levels.

Enteropathogen can be such as C.perfringens.

Performance

" animal performance " used herein by the weightening of feed efficiency and/or animal and/or can pass through feed conversion rate And/or the digestible energy in the digestibility (such as amino acid digestibility) and/or feed for passing through the nutrients in feed or metabolism The ability of the negative effect of necrotic enteritis can and/or be avoided by nitrogen reservation and/or by animal and/or is exempted from by individual Epidemic disease reacts to measure.

Preferably, " animal performance " is measured by the weightening and/or feed conversion rate of feed efficiency and/or animal.

" improved animal performance " means compared with the feed of the feed addictive composition not comprising the present invention, due to making With the feed addictive composition, the feed efficiency in individual improves and/or weightening increases and/or feed conversion rate reduces and/ Or the digestibility of the nutrients or energy in feed improves and/or nitrogen retains the negative shadow for improving and/or avoiding necrotic enteritis Loud ability improves and/or immune response improves.

Preferably, " improved animal performance " means that feed efficiency improves and/or weightening increases and/or feed conversion rate drop It is low.

The term as used herein " feed efficiency " refers to that quantity-unlimiting feeding animals or fed to it refer to during a period of time The amount of the weightening occurred in animal when quantitative food.

" feed efficiency of raising " means to feed with there is no the feed addictive composition of the present invention Animal is compared, and the weightening for causing per unit to take in feed using the feed addictive composition in feed increases.

Feed conversion rate (FCR)

The term as used herein " feed conversion rate " refers to feeding to make to measure as defined in the weight gain of animal to animal Forage volume.

Improved feed conversion rate means lower feed conversion rate.

" lower feed conversion rate " or " improved feed conversion rate " means to combine using feed addictive in feed Object causes required feeding when the weight of animals increase specified amount to be less than in the feed not comprising feed addition to the forage volume of animal Required forage volume when the weight of animals being made to increase equal amount when agent composition.

Nutrient digestibility

What Nutrient digestibility used herein meant to disappear from the specified section (such as small intestine) of gastrointestinal tract or gastrointestinal tract The score of nutrients.Nutrient digestibility can measure for be applied to individual with from individual excrement come out between difference, Or be applied to individual be retained in gastrointestinal tract specify section (such as ileum) on digest between difference.

Nutrient digestibility used herein can measure taken in nutrition by collecting total excreta during a period of time Difference between object and the nutrients drained；Or utilize and will not be absorbed by animal, and researcher is allowed to calculate entire gastrointestinal tract Or the inert labels of the amount of the nutrients to be disappeared in gastrointestinal tract section measure.This inert labels can be titanium dioxide, Chromium oxide or acid-insoluble ash.Digestibility is represented by percentage of the nutrients in feed, or is expressed as digestible battalion Support the mass unit of the nutrients in mass unit/feed of object.

Nutrient digestibility used herein covers starch digestibility, fat digestibility, protein digestibility and amino acid Digestibility.

Energy digestibility used herein means that consumed feed gross energy subtracts the gross energy of excrement, or consumed Feed gross energy subtracts the gross energy for the remaining digest that animal gastrointestinal tract is specified on section (such as ileum).Generation used herein It can refers to Apparent metabolizable energy to thank, and means that consumed feed gross energy subtracts and contain in excrement, urine and the gaseous product of digestion Gross energy.Energy digestibility and metabolic energy can measure specifies section for drained in the intake of gross energy and excrement or gastrointestinal tract Present in difference between gross energy in digest, using method identical with the digestibility of nutrients is measured, for Elimination of nitrogen carries out appropriate correction to calculate the metabolic energy of feed.

Nitrogen retains

Nitrogen used herein retains the ability for meaning individual from diet retained nitrogen as weight.It is more than daily in the excretion of nitrogen Intake when occur negative nitrogen balance and to be frequently observed the negative nitrogen balance when losing muscle.Positive nitrogen equilibrium is often given birth to muscle It is long related, it is especially related to the muscle growth in the animal just grown.

Nitrogen, which retains, to be measured as absorbed nitrogen by means of total collection of the excreta during a period of time and urine and be drained Nitrogen between difference.It is drained it should be understood that the nitrogen drained includes non-digesting protein, endogenous protein from feed Object, microprotein and urinary nitrogen.

Survival rate

The term as used herein survival rate means the number of the individual still to live.Term " improved survival rate " can be Another saying of " death rate of reduction ".

Dressing percentage and meat productivity

The term as used herein dressing percentage means after business or the slaughtering process of experiment, ratio as live weight Trunk amount.Term trunk means the animal bodies butchered by food, wherein head, internal organ, the part of four limbs and feather or skin Skin has removed.The term as used herein meat productivity means the amount of the edible meat of the ratio as live weight, or as live body The amount of the specified meat of the cutting of the ratio of weight.

Weightening

The present invention also provides the methods for increasing the weightening in individual (such as poultry or pig) comprising gives individual feeding packet The feed of feed addictive composition containing the present invention.

" increased weightening " refers to compared with the animal for the feed that feed addictive composition is not present with feeding, is wrapped in feeding Animal has increased weight when feed containing the feed addictive composition.

Increased weightening can be at least 0.5%, at least 1%, at least 2%, at least 3%, at least 5%, at least 10%, extremely Few 12%, at least 15%, at least 20% or at least 30%.In some embodiments, which can be at least 50% or extremely Few 100%.

The increase of weightening can be that just in contrast, in the control, used feed does not include DMF and high dose (i.e.>1500FTU/kg feeds) phytase.On the other hand, the increase of weightening can be just comprising another inositol six For the use of the feed of phosphatase, wherein the phytase with low dosage (such as<1000FTU/kg feeds) it uses.

Clinical or subclinical enteropathy attack

Term " clinical disease " means that anatomy to be enough to generate identifiable disease indication and symptom or physiology change Become the stage of the course of the pathological conditions started.

Term " subclinical disease " means or is related to noticing the course of disease stage before symptom for the first time.

Enteropathy can be the arbitrary intestines infection that individual can be suffered from.

In an example, enteropathy can be caused by genic organisms (such as pathogenic bacteria).

In one embodiment, enteropathy can be caused by one or more in following organism：Pathogenic fusobacterium Species, such as C.perfringens and/or clostridium difficile and/or Escherichia coli and/or Salmonella ssp and/or bent stick Ella species.

In one embodiment, enteropathy can be caused by one or more in following pathogen：C.perfringens And/or clostridium difficile and/or Escherichia coli, preferably C.perfringens and/or clostridium difficile, more preferable C.perfringens.

Necrotic enteritis

Necrotic enteritis is the acute or chronic enterotoxemia observed in chicken, turkey and duck in world wide, by C.perfringens causes.Necrotic enteritis is often characterized as the cellulosic necrotic enteritis of typically mid-small bowel.The death rate Can be 5% to 50%, normally about 10%.Infection broadcasts life by excrement-oral instructions.The spore resistance of genic organisms is high.It lures Cause of disease element includes globidiosis, diet (high protein), may attach most importance in duck stress, high viscosity diet (include often with diet A large amount of rye is related to wheat), the feed that pollutes and/or water, other wasting diseases.

The present invention relates to increase method of the individual to the resistance of necrotic enteritis.In other words, the present invention relates to avoid or subtract The negative effect of light necrotic enteritis.

The term as used herein " resistance " can cover term " tolerance ".Therefore, in one embodiment, individual can be with Necrotic enteritis can be resistant to necrotic enteritis non-resistant but individual, i.e., to the performance of individual without negative effect.

In one embodiment, it the present invention relates to the feed addictive composition of the present invention, is used to control in individual Treat or prevent necrotic enteritis.In general, individual can be by or will be by C.perfringens and/or eimeria (Eimeria) individual of species attack.This attack may be from environment, or apply viable microbial, example in feed or drinking water Such as when using coccidial vaccine living.

In another embodiment, the present invention relates to feed addictive composition, be used in individual prevent and/or Treat globidiosis.

The method that the present invention furthermore provides prevention and/or treats necrotic enteritis and/or globidiosis, wherein to individual Using the feed addictive composition of a effective amount of present invention.

Immune response

Immune response used herein means that DFM adjusts one of multiple modes of immune system of animal, including increases Antibody generates, raises cell-mediated immune, up-regulation proinflammatory cytokine and increase toll sample receptor signal grantings.It should be understood that DFM can be conducive to the immunostimulation of gastrointestinal tract host is protected to resist the disease, and can be right to the immunosupress of gastrointestinal tract Host is advantageous, this is because less nutrients and energy be used to support immune function.

Preferably, immune response is cell immune response.

Preferably, immune labeled immune response is measured by observing.

Pathogenic bacteria

The term as used herein pathogenic bacteria mean such as toxigenicity clostridium species, such as C.perfringens and/or large intestine Bacillus and/or Salmonella ssp and/or campylobacter species.In one embodiment, pathogenic bacteria can be fowl cause Characteristic of disease species Escherichia coli.

The present invention can reduce the pathogenic bacteria population in individual gastrointestinal tract.

Nutrients is drained

In one embodiment, the present invention relates to the nutrients reduced in muck excretions.This is to reducing environmental hazard tool There is positive effect.For example, in preferred embodiments, the present invention relates to the nitrogen and/or phosphorus content that reduce in individual muck.Cause This, this can reduce the amount of nitrogen and/or phosphorus in environment, this can be advantageous.

Probiotics (probiotic)

For some applications, it is believed that the DFM in the present composition can play probiotic bacterial cultures effect.To the present invention's Composition adds other probiotics and/or probiotics are also within the scope of the invention.

Here, probiotics is：

" non-digestible food ingredients, by selective stimulating is a kind of or the growth of the beneficial bacteria of limited quantity and/or Activity and advantageously influence host ".

The term as used herein " probiotic bacterial cultures " definition viable microbial (e.g., including bacterium or yeast), in example Can advantageously influence host organisms when such as applying with sufficient amount intake or part, i.e., by assign host organisms it is a kind of or A variety of verifiable health benefits.Probiotics can improve the microbial balance of one or more mucomembranous surfaces.For example, mucous membrane table Face can be intestines, urethra, respiratory tract or skin.The term as used herein " probiotics ", which is also covered by, can stimulate the beneficial of immune system to divide Branch and the viable microbial for reducing the inflammatory reaction in mucomembranous surface (such as intestines) simultaneously.

Although for probiotics absorbs, lower limit or the upper limit is not present, it has been proposed that at least 10⁶-10¹², preferably at least 10⁶-10¹⁰, preferably 10⁸-10⁹Cfu will be effective to reaching beneficial health effect in individual as daily dose.

Separation

On the one hand, expediently, enzyme used in the present invention or DFM can be the forms of separation.Term " separation " is anticipated Refer to enzyme or DFM least substantially at without the enzyme or DFM in nature it is natural associated or see in nature at least one Kind other compositions.The enzyme or DFM of the present invention can be substantially free of the originally associated one or more pollutants of the substance Form provide.Thus, for example, it can be substantially free of the polypeptide of one or more potential pollutions and/or nucleic acid molecules.

Purifying

On the one hand, it is preferable that enzyme and/or DFM of the invention is the form of purifying.Term " purifying " mean enzyme and/ Or DFM exists with high level.Enzyme and/or DFM advantageously main component present in composition.Preferably, it is at least about 90%, or at least about 95% or at least about 98% horizontal exists, the level be relative to the total composition considered with dry weight/ It is measured based on dry weight.

Imagine within the scope of the invention, embodiment of the present invention can be combined so that arbitrary characteristics as described herein Combination be included within the scope of the present invention.Specifically, imagining within the scope of the invention, bacterium can be showed simultaneously Any one of therapeutic effect.

With reference to the following drawings and embodiment, only description description is of the invention by way of example.

Embodiment

Embodiment 1

Method

It is bought from commercial hatchery and amounts to 1600 1 age in days Cobb male chick.When studying beginning, subregion (block) 50 male chick are distributed to each processing henhouse (pen).Research is made of (table 1) following processing：

The experimental design of 1. embodiment 1 of table

Different diet	C.perfringens is attacked	Phyzyme XP¹	DFM (CFU/g feeds)²
				1	It is no	500FTU/kg	Nothing
2	It is	500FTU/kg	Nothing
				3	It is	500FTU/kg	Enviva Pro(7.5x 10⁴CFU/g)
4	It is	3000FTU/kg	Enviva Pro(7.5x 10⁴CFU/g)

¹Phyzyme XP are the 6-phytases from Escherichia coli provided by Danisco A/S BJ Rep Office.

²EnvivaBe bacillus subtilis strain Bs2084, the LSSAO1 provided by Danisco A/S BJ Rep Office and The combination of 15AP4.

Start in research, the 23rd day, the 35th day and terminate to record the chicken weight of each henhouse when (the 42nd day).Henhouse is to survey Measure unit.The feeding dorking diet in the form of clast (opened guideway) or pellet (growth material and fattening are expected).Diet meets or more than NRC Standard (table 2).Mixer is rinsed to prevent diet cross contamination.All processing feeds are mixed with Davis S-20 mixers to be used in combination California pellet mills pill (cold pellet temperature is 65 DEG C to 70 DEG C).Since each batch, it is intermediate and terminate from every One processing diet is collected sample and is blended together to confirm enzymatic activity and the Enviva Pro presence in feed.

The experiment diet of 2. embodiment 1 of table forms

Chicken quantity-unlimiting feed received suitable for the processing from the 0th to 42 day.Enzyme and Enviva Pro are by Danisco to be suitble to Mixture for all experiment process and horizontal offer.Henhouse, which is arranged in facility, to be prevented from being in direct contact, to avoid pollution. Growth material is changed to by opened guideway at the 23rd day.At the 35th day growth material was substituted with fattening material.When changing feed every time, subregion Feeder is pipetted from henhouse, is weighed again, empty and reloads processing diet appropriate.Research last day to feed It weighs.Daily henhouse is checked for the death rate.When chicken is eliminated or finds death, record date and removal weight (Kg).Gross necropsy is implemented to determine gender and the possible cause of death to all dead or superseded chicken.Pay attention to gangrenosum acne intestines Scorching sign.

All henhouses all have about 4 inch straw in a mattress, have one layer of fresh pine bits.

Spraying vaccine is carried out to it with business coccidiosis vaccine (Coccivac-B) before all chickens are positioned over henhouse Inoculation.At the 20th, 21 and 22 day, in addition to processing 1, all chickens are given with the broth culture of C.perfringens.Using known The field separation strains of NE and the C.perfringens from commercial meat chickens operation can be caused as challenge organism.Daily using new Fresh inoculum.Titer level is about 1.0 × 10^8-9.Each henhouse receives same amount of inoculum.It is raised by being mixed into tubular type It feeds and applies inoculum in feed present in device base portion.At the 23rd day, 5 chickens are selected from each henhouse, implement euthanasia, point Group is weighed, and check necrotic enteritis damage there are degree.Scoring is based on 0 to 3 point, wherein 0 is normal, and 3 be most serious (0=is without 1=is slight, 2=moderates, and 3=is notable/serious；Hofacre etc., 2003J.Appl.Poult.Res.12:60–64). Adjoint medicinal treatment is not used during research.

Average value is detached with paired t-test.Think in P<0.05 time with significant difference.It is single using henhouse as experiment Member.

As a result

From the 23rd day up to off-test as shown in (Fig. 1), C.perfringens attack substantially reduce weight gain.It mends It fills Enviva Pro and significantly improves the weight gain of each sample day to the level of positive control.But further supplement high flesh Alcohol six-phosphatase dosage (processing 10) improve weight to the 12nd, 23 and 35 day the level for being significantly higher than positive control and the 42nd day Numerically bigger level.

As from the 23rd day shown in the off-test (Fig. 2), C.perfringens attack significantly improves feed conversion rate (FCR).Enviva Pro are added into diet causes FCR to be significantly reduced from negative control.But supplement high phytase Dosage reduces FCR to the identical level with seen in positive control.

Supplement Enviva Pro make the death rate observed reduce (Fig. 3), supplement high phytic acid enzyme dosage make its into One step numerically reduces.This severity that can be construed to C.perfringens attack reduces, this is high six phosphorus of inositol of supplement The seen Injury score numerically reduced supports (Fig. 4) when sour enzyme dosage, which is markedly different from the damage of positive control Wound scoring.

With using Enviva Pro high phytic acid is utilized compared with the phytase of standard (500FTU) dosage Enzyme dosage (3000FTU/kg) and the combination of Enviva Pro have apparent benefit, this is the FCR reduced and increased weightening It is proved.These benefits are construed to reduced damage of intestines.

Embodiment 2

Material and method

The male chick of 1040 1 age in days Cobb x Cobb 500 is amounted to from commercial hatchery purchase.When studying beginning, Subregion is randomly assigned 10 chick to Petersime chicken coops by each processing.Only selection healthy chicken is for testing, and is entirely studying Do not replace chicken in the process.Research is made of (table 1) following processing：

The experimental design of 1. embodiment 2 of table

¹Enviva Pro are the combination of bacillus subtilis strain Bs2084, LSSAO1 and 15AP4, by Danisco public affairs Department provides, and is added by 75,000CFU/g feeds.

²AxtraPhy^TMIt is the 6-phytase from Buttiauxella, is provided by Danisco A/S BJ Rep Office.

Chicken weight is recorded when research starts (the 0th day) and the 13rd day, the 21st day and research terminates (the 28th day).Chicken coop is Measuring unit.Diet is fed in the form of thickener, and is formulated as meeting or more than NRC standards (table 2).Mixer is rinsed to prevent day Cross contamination between grain.Since each batch, it is intermediate and terminate from each processing diet collection sample and mix use In analyzing enzymatic activity in feed and DFM exists.

The experimental diets of 2. embodiment 2 of table form

All chicken feeding " business style " daily rations are until the 9th day；From the 10th day, feeding processing daily ration.Changing feed When, feeder is pipetted from henhouse, is weighed again, empties and reloads processing diet appropriate.In the last day pair of research Feed is weighed.Daily henhouse is checked for the death rate.When chicken is eliminated or finds death, record date and removal weight (Kg).Gross necropsy is implemented with the possible cause of death of determination to all dead or superseded chicken.Record the mark of necrotic enteritis As.Chicken can get the quantity-unlimiting feed of proper treatment during entire research.

Disease induction is carried out according to study site Standard Operating Procedure (SOP).In short, at the 13rd day, all chickens are all used Eimeria maxima (Eimeria maxima) oral vaccination, inoculum contain every chicken about 5,000 egg capsule.At the 18th day, In addition to processing 1 and 3, all chickens all receive the broth culture of C.perfringens, which contains about 10⁸cfu/ml.Institute There is processing henhouse to receive same amount of inoculum.By fresh to be administered once a day in the feed that is mixed into feeder base portion C.perfringens inoculum, using three days (the 18th, 19 and 20 day).At the 21st day, 3 chickens are selected from each henhouse, are implemented Euthanasia, grouping weighs, and check necrotic enteritis damage there are degree.Scoring is based on 0 to 3 point, wherein 0 be it is normal, and 3 It is that (0=is without 1=is slight, 2=moderates, and 3=is notable/serious for most serious；Hofacre etc., 2003J.Appl.Poult.Res.12:60–64).Adjoint medicinal treatment is not used during research.

For showing data, using JMP softwares, the effect of processing is examined with ANOVA, is used in combination paired t-test separation average Value.Think in P<0.05 time with significant difference.Using chicken coop as experimental considerations unit.For pH and organ weight data, every Chicken is experimental considerations unit.

In order to examine interaction, main effect and their phase including phytase and Enviva Pro are used The factorial arrangement of interaction, with the subgroup of ANOVA analyzing processings (processing 4 to 13).

As a result

Compared with control not under fire, weightening (BWG) and feed conversion rate (FCR) with regard to the 21st day and the 28th day and Speech, C.perfringens significantly reduce the performance (for T1 to T2, T3 is to T4) (table 3) of chicken.Lacking C.perfringens attack In the case of, the reduction of diet P does not influence the BWG of the 21st day or the 28th day.But it is attacking there are C.perfringens In the case of, there are the reductions of significant numerical value between high P diets and low P diets by BWG.These observation results are consistent with the data of FCR.

In individually supplement phytase, there is variable effect to the performance of chicken；But in all cases, There are numerical value to improve to the horizontal level higher than low P diets (T4) under fire by 21 days BWG.It is being obtained the results show that Under conditions of necrotic enteritis attack, the performance of chicken is not further improved in high phytic acid enzyme dosage；Because in 10,000FTU The minimum BWG only with phytase treatment is found under the phytase (T8) of dosage, this is markedly different under fire Low P diets (T4).Equally, at the 28th day, in individually supplement phytase, BWG is in the inositol six higher than 1500FTU There are numerical value reductions under phosphatase level.

In individually supplement Enviva Pro (T9), the 21st day BWG is significantly improved from low P diets (T4) under fire, and The 28th day further improve to low level of the P diets without significant difference not under fire.For FCR, Enviva Pro There is the significant decrease from control under fire in supplement, but be not enough to the negative effect completely to attack resistance.

When combination, compared with when individually supplementing phytase, the 21st day and the 28th day, six phosphorus of inositol of high dose Sour enzyme and Enviva Pro lead to more uniform reaction for BWG.For FCR, which existed at the 21st day (Fig. 5) Synergistic effect.In individually supplement phytase, there is performance at 500,1500 (T7) and 3000FTU/kg levels Improve (reduction of FCR), and under conditions of necrotic enteritis attack more than phytic acid enzyme dosage when there are the drops of performance It is low.But in combination be added probiotics when, with only at most and include 3000FTU horizontal processing (T12) compared with, FCR mono- Causing ground, there are numerical value reductions.But at 10,000FTU (T13), there are big to be substantially reduced, and causes in all processing groups Minimum FCR.In specific effect (the excluding positive control T1, T2 and T3) for investigating combination, there is combination to the 21st day FCR Significantly affect (P=0.0319) (table 4).

The reduction of diet AvP causes substantially reducing (table 3) for shin bone bone ash, shin bone bone ash to be the instructions of phosphorus bioavailability Object.C.perfringens attack causes the small numerical value of tibial bone ash content to reduce, this may be that the intestines caused by infection damage Wound reduces the ability that intestines effectively absorb nutrients.Individually supplement phytase cause shin bone bone ash % restore to not by High level of the P diets (T1) without significant difference of attack.In individually supplement Enviva Pro (T9), shin bone bone ash % numerical value increases The level higher than low P diets (T3) not under fire is added to, this demonstrate that the product fights production for the reduction of shin bone bone ash The ability of the influence of gas capsular clostridium attack, this may indicate that the recovery of gut function.When supplementing the combination, exist to inositol six The dose response of phosphatase reaches the level higher than high P diets (T1) not under fire at 3000FTU, in individually supplement flesh This improvement is not observed when alcohol six-phosphatase.

The processing of table 3. is to performance parameter, feed intake, weightening, the influence of feed conversion rate and the 21st day bone ash data

The combination of 4. phytase+Enviva Pro of table is significantly affected on the 21st day FCR's

C.perfringens attack, which has the death rate and Injury score, to be significantly affected (table 4).It is under fire not dead in group The shortage of rate and the shortage of Injury score show under fire group and under fire there is no pollutions between group.With regard to Injury score Speech is all not present under arbitrary phytase level between phytase and EnvivaPro+ phytases and shows Write difference (T4-8 is to T9-13).But 1500 (T6) and 3000FTU (T7) are handled, only using phytase treatment Low P diets under fire level on there are the increases of the numerical value of the death rate.The addition of EnvivaPro cause the death rate from by Attack low P diets significantly reduce, with only with phytase treatment compared with, also reduce the phytase agent of combination The variation of the death rate between amount.

Influence of the processing of table 5. to the death rate (%) and necrotic enteritis (NE) Injury score

Embodiment 3

Method

It is bought from commercial hatchery and amounts to 1080 1 Japanese instar chicklings.When studying beginning, subregion divides at random to each henhouse With 15 chick.Research forms (table 1) by following processing, each processing has 8 repetition henhouses.

Table 1：Experimental design

¹EnvivaIt is the combination of bacillus subtilis strain Bs2084, LSSAO1 and 15AP4, by Danisco Company provides.

²Axtra Phy are six phosphorus of 6- inositols from Buttiauxella that can be obtained from Danisco Animal nutrition company Sour enzyme.

Research start (the 0th day), the 6th day, the 21st day and research terminate (the 35th day) when record chicken weight.Henhouse is to survey Measure unit.It prepares diet and instructs (table 2) to meet NRC.All processing feeds use Davis S-20 mixers to mix.In diet Between rinse mixer to prevent the cross contamination of diet.Since each batch, it is intermediate and terminate from each processing diet Sample, mixing are collected, and is analyzed, to confirm enzymatic activity and the DFM presence in feed.Diet is fed in the form of thickener.

All henhouses all have about 4 inch straw in a mattress, have one layer of fresh pine bits.All chickens are being put into henhouse prosopyle Clothes receive business coccidiosis vaccine.

Table 2：Experimental diets form

All chickens fed business style opened guideway daily ration from the 0-6 days.From the 7th day, growth material of the feeding suitable for processing Diet is until the 21st day.Terminated fattening material diet of the feeding in (the 35th day) suitable for processing from the 21st day to research.Changing feed When, feeder is pipetted from henhouse, is weighed again, empties and reloads processing diet appropriate.In the last day pair of research Feed is weighed.Daily henhouse is checked for the death rate.When chicken is eliminated or finds death, record date and removal weight (Kg)。

For showing data, average value is detached with paired t-test.Think in P<0.05 time with significant difference.Use chicken House is used as experimental considerations unit.

As a result

Feed conversion rate (FCR) significantly improves when individually the dosage of increase phytase does not result in 35 age in days.Institute There are addition Enviva Pro under phytic acid enzyme level to reduce FCR.As phytic acid enzyme dosage increases, from addition The benefit of Enviva Pro also increases.Horizontal phytase generation is contained up to contain and the diet without Enviva Pro Between FCR maximum difference.

Table 3：Performance results

Under higher phytic acid enzyme dosage, compared with when individually supplementing phytase, Enviva is supplemented The numerical value that there is weightening (BWG) when Pro increases (table 3).It is not observed under minimum phytic acid enzyme dosage (500FTU) This effect.

Embodiment 4

Material and method

1 age in days Cobb male chick is bought from commercial hatchery.When studying beginning, subregion is randomly assigned to each chicken coop 10 males.Each processing has 8 repetition chicken coops.Research is made of (table 1) following processing.

Table 1：The experimental design of embodiment 4

¹HiPhos is Ronozyme HiPhos, including the phytase from Bu Shi citric acid bacteriums, it can be from Supreme Being Si Man/Novozymes Company obtains.

²EnvivaIt is the combination of bacillus subtilis strain Bs2084, LSSAO1 and 15AP4, by Danisco Company provides.

Chicken weight is recorded when research starts (the 0th day), the 21st day and research terminate (the 28th day).Chicken coop is to measure list Position.Diet meets or more than NRC standards (table 2).All processing feeds use Davis S-20 mixers to mix.Between diet Mixer is rinsed to prevent the cross contamination of diet.Since each batch, it is intermediate and terminate from the collection of each processing diet Sample is simultaneously blended together, to confirm enzymatic activity and the Enviva Pro presence in feed.

Table 2：The experimental diets of embodiment 4 form

Daily ration is until the 9th day before all chicken feeding business opened guideways；From the 10th day, feeding processing daily ration.It is raised changing When material, feeder is pipetted from chicken coop, is weighed again, empty and reloads processing diet appropriate.In the last day of research It weighs to feed.Daily chicken coop is checked for the death rate.When chicken is eliminated or finds death, record date and removal weight It measures (Kg).Gross necropsy is carried out with the possible cause of death of determination to all dead or superseded chicken.Record necrotic enteritis Sign.Chicken can get the quantity-unlimiting feed of proper treatment during entire research.

Disease induction is carried out according to study site Standard Operating Procedure (SOP).In short, at the 13rd day, all chickens are all used Eimeria maxima oral vaccination, inoculum contain every chicken about 5,000 egg capsule.At the 18th day, in addition to processing 1 and 3, own Chicken all receives the broth culture of C.perfringens, which contains about 10⁸cfu/ml.All processing henhouses receive identical The inoculum of amount.By being administered once a day fresh C.perfringens inoculation in the feed that is mixed into feeder base portion Object, using three days (the 18th, 19 and 20 day).At the 21st day, 3 chickens being selected from each henhouse, implement euthanasia, grouping is weighed, And check necrotic enteritis damage there are degree.Scoring is based on 0 to 3 point, wherein 0 is normal, and 3 be that (0=is without 1 for most serious =slight, 2=moderates, 3=are notable/serious；Hofacre etc., 2003J.Appl.Poult.Res.12:60–64).In the research phase Between do not use adjoint medicinal treatment.

At the 21st day, each chicken coop measured the pH of the ileum digest of 2 chickens.It is visited by the way that Sensorex lances are punctured pH Needle is inserted into each segment to measure the pH of digest.In two chickens, gizzard is also taken out, and weigh.At the 21st day, two chickens are taken out Right shin bone, be dried overnight and weigh at 100 DEG C；Then fat-extraction, and 600 DEG C of burnings in Muffle furnace are carried out to sample 16 hours, to measure not fatty bone ash.Then by percentage bone ash be calculated as remaining bone ash tumour with it is dry fat-free The ratio of bone weight is multiplied by 100.

Statistical analysis

For showing data, average value is detached with paired t-test.Think in P<0.05 time with significant difference.Use chicken Cage is as experimental considerations unit.

As a result

In the case where necrotic enteritis is attacked, increasing the dosage of HiPhos causes 21 age in days feed conversion rates to improve (figure 7；The more feeds of per unit weightening).Enviva Pro, which are added, under all levels comprising phytase causes to reduce (improved FCR).In the case where containing up to horizontal phytase (3000FTU/kg), the addition of Enviva Pro gradually subtracts The weak FCR arrived in 1500FTU/kg observed at doses is improved.

Table 3：Performance results from embodiment 4

Effect of gain of the Enviva Pro on phytase is in high phytic acid enzyme dosage (3000FTU/kg) Under significantly, wherein compared with control treatment not under fire, increase weight without significant difference.

Influence of the phytase to shin bone bone ash percentage under fire compares no significant difference with feminine gender, shows even At high doses, phytase does not make significant difference (Fig. 8) to the shin bone bone ash in the case of attack yet.But such as institute in table 4 See, the addition of Enviva Pro significantly improves shin bone bone ash percentage, makes it close in highest phytic acid enzyme dosage The percentage observed in positive not under fire control under (3000FTU/kg).

Table 4：Other electrophysiology measurements from embodiment 5

The ileum pH that high-level phytase (3000FTU/kg) is improved the 21st day is more than related control, and Enviva The addition of Pro makes it reduce, and with Enviva Pro, individually minimum pH is observed in processing.In under fire negative control and maximum dose level Hi Phos (3000FTU/kg) in observe minimum gizzard weight.The addition of Enviva Pro is under 3000FTU/kg dosage Gizzard weight is numerically increased, but not significantly.

Attack significantly improves the death rate, and the addition of phytase reduces the death rate.Combination with Enviva Pro into One step reduces the death rate.

All publications mentioned in description above are hereby incorporated by reference.To those skilled in the art, it shows And be clear to, described the method for the present invention and system can be made without departing substantially from scope and spirit of the present invention Numerous modifications and variations.Although specific preferred embodiment has been combined to describe the present invention, it should be appreciated that claimed Invention should not undeservedly be limited to this kind of specific embodiment.In fact, the mode for carrying out the present invention to biology Obvious a variety of modifications are intended in following following claims for chemistry and biotechnology or various equivalent modifications Within the scope of.

Claims

1. the performance for improving individual, or the digestibility for improving the raw material in feed, or retain for improving nitrogen, or use It absorbs and retains in improving diet phosphorus, or the effect of for improving phytase, or for improving feed conversion rate (FCR), or for improving the weightening in individual, or for improving the feed efficiency in individual, or for adjusting the immune of individual Reaction, or the population for reducing the pathogenic bacteria in individual gastrointestinal tract, or the side for reducing the excretion of the nutrients in muck Method, the method includes the combinations at least one Direct-Fed Microbials (DFM) and phytase of individual application, wherein The phytase is applied to the individual by the dosage more than 1500FTU/kg feeds.

2. the purposes of the combination of at least one Direct-Fed Microbials and phytase, wherein phytase are by being more than The dosage of 1500FTU/kg feeds uses, the performance for improving individual, or the digestibility for improving the raw material in feed, or Retain for improving nitrogen, or absorb and retain for improving diet phosphorus, or the effect of for improving phytase, or is used for Improve feed conversion rate (FCR), or for improving the weightening in individual, or for improving the feed efficiency in individual, or is used for The immune response of individual, or the population for reducing the pathogenic bacteria in individual gastrointestinal tract are adjusted, or for reducing in muck Nutrients is drained.

3. the method or purposes of claim 1 or claim 2, wherein Direct-Fed Microbials are that antipathogen Direct-fed is micro- Biology.

4. the method or purposes of claim 1 or claim 2, wherein Direct-Fed Microbials are bacteriums living.

5. the method or purposes of claim 1 or claim 2, wherein pressing more than the dosage of 2000FTU/kg feeds to described Body applies the phytase.

6. the method or purposes of claim 1 or claim 2, wherein pressing more than the dosage of 3000FTU/kg feeds to described Body applies the phytase.

7. the method or purposes of claim 1 or claim 2, wherein composition include at least three kinds of Direct-Fed Microbials.

8. the method or purposes of claim 1 or claim 2, wherein Direct-Fed Microbials are to inhibit the intestines of pathogenic microorganisms The bacterial strain inside colonized.

9. the method or purposes of claim 8, wherein the pathogenic microorganisms is C.perfringens or Escherichia coli, or both Combination.

10. the method or purposes of claim 1 or claim 2, wherein Direct-Fed Microbials include in subordinate One or more bacteriums：Bacillus, enterococcus spp, Pediococcus, saccharomyces, Bifidobacterium, Propionibacterium, breast Bacillus, lactococcus, aspergillus and combinations thereof.

11. the method or purposes of claim 1 or claim 2, wherein Direct-Fed Microbials include in following species One or more bacteriums：Bacillus subtilis, bacillus licheniformis, bacillus amyloliquefaciens, Bacillus cercus, dung intestines Coccus, Pediococcus acidilactici, saccharomyces cerevisiae, animal bifidobacteria animal subspecies, lactobacillus reuteri, Lactobacillus rhamnosus, acidophilus breast Bacillus, Lactobacillus brevis, Lactobacillus salivarius subspecies, Lactobacillus farciminis, Lactococcus lactis, Lactobacillus casei, Lactobacillus salivarius, Lactobacillus rhamnosus, clostridium butyricum, aspergillus oryzae, production propionibacterium acide-propionici, propionibacterium jensenii, enterococcus faecium, Pediococcus acidilactici And combinations thereof.

12. the method or purposes of claim 1 or claim 2, wherein Direct-Fed Microbials are one in following bacterial strain Or it is multiple：Bacillus subtilis BS18 (NRRL B-50633), bacillus subtilis BS278 (NRRL 50634), withered grass gemma Bacillus 4-7d (NRRL B-50505), bacillus subtilis 3-5h (NRRL B-50507), bacillus subtilis AGTP BS3BP5 (NRRL B-50510), bacillus subtilis BS918 (NRRL B-50508), bacillus subtilis AGTP BS1013 (NRRL- 50509), bacillus subtilis AGTP 944 (NRRL B-50548), bacillus subtilis AGTP BS442 (NRRL B- 50542), bacillus subtilis AGTP BS1069 (NRRL B-50544), bacillus subtilis AGTP BS521 (NRRL B- 50545), bacillus subtilis BS2084 (NRRL B-50013), bacillus subtilis LSSA01 (NRRL B-50104), withered grass Bacillus B27 (NRRL B-50105), bacillus subtilis 3A-P4 (PTA-6506), bacillus subtilis 15A-P4 (PTA- 6507), bacillus subtilis 22C-P1 (PTA-6508), bacillus subtilis BL21 (NRRL B-50134), lichens gemma bar Bacterium BL21 (NRRL B-50134), bacillus licheniformis 3-12a (NRRL B-50504), bacillus licheniformis 4-2a (NRRL B- 50506), bacillus licheniformis 842 (NRRL B-50516), production propionibacterium acide-propionici P261 (NRRL B-50131), production propionic acid Propionibacterium P179 (NRRL B-50133), production propionibacterium acide-propionici P169 (PTA 5271), production propionibacterium acide-propionici P170 (PTA 5272), propionibacterium jensenii P63 (NRRL B-30979), propionibacterium jensenii P195 (NRRL B-50132), lactic acid It is galactococcus ID7 (PTA 6103), Lactococcus lactis JD19 (PTA 6104), lactobacillus acidophilus A2020 (NRRL B-30977), thermophilic Lactobacillus lactis A4000h (NRRL B-30978), lactobacillus acidophilus PIBc6 (NRRL B-50103), Lactobacillus brevis LBR 1000 (NRRL B-30982), Lactobacillus casei LC222 (NRRL B-30983), family name's lactobacillus PLCB6 (NRRL B-50518), saliva It is lactobacillus o246i33w (NRRL B-50102), Lactobacillus brevis AJ25 (PTA-6099), Lactobacillus brevis HE17 (PTA-6100), short Lactobacillus 1E-1 (PTA-6509), lactobacillus lactis Cl15 (PTA-6101), lactobacillus lactis DJ6 (PTA-6102), rhamnose Lactobacillus (CNCM I-3698), Lactobacillus farciminis (CNCM I-3699), enterococcus faecium EF141 (EN-1) (NRRL B- 30981), enterococcus faecium 2-1d (NRRL B-50519), Pediococcus acidilactici PIJe3 (NRRL B-50101), Pediococcus acidilactici O246e42 (NRRL B-50171) and combinations thereof.

13. the method or purposes of claim 1 or claim 2, wherein phytase are 6-phytase or 3- Phytase.

14. the method or purposes of claim 13, wherein phytase are 6-phytases.

15. the method or purposes of claim 1 or claim 2, wherein phytase are Escherichia coli phytic acids Enzyme or Buttiauxella phytase or Citrobacter phytase or Hafnia phytic acid Enzyme or aspergillus phytase or Penicillium phytase or trichoderma phytase or Escherichia coli Phytase or Hansenula phytase or Peniophora category (Peniphora) phytase.

16. the method or purposes of claim 1 or claim 2, wherein DFM presses 3.75x10⁷CFU/g feeds are to 1x10¹¹CFU/ The dosage of g feeds exists.

17. the method or purposes of claim 1 or claim 2, wherein there is clinical or subclinical enteropathy attack in individual.

18. the method or purposes of claim 17, wherein clinical or subclinical enteropathy attack is caused by pathogenic bacteria.

19. the method or purposes of claim 18, wherein the pathogenic bacteria are C.perfringens or Escherichia coli.

20. the method or purposes of claim 1 or claim 2, wherein the digestibility is Nutrient digestibility, the adjusting The immune response of individual is to improve the immune response of individual.

21. the method or purposes of claim 20, wherein the Nutrient digestibility is amino acid digestibility.

22. feed addictive composition, it includes the combinations of at least one Direct-Fed Microbials and phytase, wherein Phytase is by existing below in the feed addictive composition：

A. when being added in feed by least 50g/ tonnes of (MT) feed, by 30,000FTU/g compositions or more；

B. when being added in feed by least 75g/ tonnes of (MT) feed, by 20,000FTU/g compositions or more；

C. when being added in feed by least 100g/ tonnes of (MT) feed, by 15,000FTU/g compositions or more；

D. when being added in feed by least 100g/ tonnes of (MT) feed, by 15,000FTU/g compositions or more；

E. when being added in feed by least 150g/ tonnes of (MT) feed, by 10,000FTU/g compositions or more；

F. when being added in feed by least 200g/ tonnes of (MT) feed, by 7,500FTU/g compositions or more；

G. when being added in feed by least 300g/ tonnes of (MT) feed, by 5,000FTU/g compositions or more；

And wherein Direct-Fed Microbials are pressed from 2.5x10³CFU DFM:1FTU enzymes are to 6.7x10⁶CFU:The range of 1FTU enzymes exists In the feed addictive composition.

23. the feed addictive composition of claim 22, wherein Direct-Fed Microbials are pressed from 3.8x10³CFU DFM:1FTU Enzyme is to 2.0x10⁵CFU:The range of 1FTU enzymes is present in the feed addictive composition.

24. the feed addictive composition of claim 22 or claim 23, wherein Direct-Fed Microbials are antipathogens Direct-Fed Microbials.

25. the feed addictive composition of claim 22, wherein Direct-Fed Microbials are bacteriums living.

26. the feed addictive composition of claim 22, wherein composition include at least three kinds of Direct-Fed Microbials.

27. the feed addictive composition of claim 22, wherein Direct-Fed Microbials are the enterals for inhibiting pathogenic microorganisms The bacterial strain colonized.

28. the feed addictive composition of claim 27, wherein pathogenic microorganisms are C.perfringens, Escherichia coli, sand Door Salmonella species or campylobacter species, or combinations thereof.

29. the feed addictive composition of claim 22, wherein Direct-Fed Microbials include from one in subordinate Or multiple bacterium：Bacillus, enterococcus spp, Pediococcus, saccharomyces, Bifidobacterium, Propionibacterium, lactobacillus Category, lactococcus, aspergillus and combinations thereof.

30. the feed addictive composition of claim 22, wherein Direct-Fed Microbials include one in following species A or multiple bacterium：Bacillus subtilis, bacillus licheniformis, bacillus amyloliquefaciens, Bacillus cercus, dung intestines ball Bacterium, Pediococcus acidilactici, saccharomyces cerevisiae, animal bifidobacteria animal subspecies, lactobacillus reuteri, Lactobacillus rhamnosus, acidophilus breast bar Bacterium, Lactobacillus brevis, Lactobacillus salivarius subspecies, Lactobacillus farciminis, Lactococcus lactis, Lactobacillus casei, Lactobacillus salivarius, mouse Lee's sugar lactobacillus, clostridium butyricum, aspergillus oryzae, production propionibacterium acide-propionici, propionibacterium jensenii, enterococcus faecium, Pediococcus acidilactici and A combination thereof.

31. the feed addictive composition of claim 22, wherein Direct-Fed Microbials are one or more in following bacterial strain It is a：Bacillus subtilis BS18 (NRRL B-50633), bacillus subtilis BS278 (NRRL 50634), bacillus subtilis 4-7d (NRRL B-50505), bacillus subtilis 3-5h (NRRL B-50507), bacillus subtilis AGTP BS3BP5 (NRRL B-50510), bacillus subtilis BS918 (NRRL B-50508), bacillus subtilis AGTP BS1013 (NRRL- 50509), bacillus subtilis AGTP 944 (NRRL B-50548), bacillus subtilis AGTP BS442 (NRRL B- 50542), bacillus subtilis AGTP BS1069 (NRRL B-50544), bacillus subtilis AGTP BS521 (NRRL B- 50545), bacillus subtilis BS2084 (NRRL B-50013), bacillus subtilis LSSA01 (NRRL B-50104), withered grass Bacillus B27 (NRRL B-50105), bacillus subtilis 3A-P4 (PTA-6506), bacillus subtilis 15A-P4 (PTA- 6507), bacillus subtilis 22C-P1 (PTA-6508), bacillus subtilis BL21 (NRRL B-50134), lichens gemma bar Bacterium BL21 (NRRL B-50134), bacillus licheniformis 3-12a (NRRL B-50504), bacillus licheniformis 4-2a (NRRL B- 50506), bacillus licheniformis 842 (NRRL B-50516), production propionibacterium acide-propionici P261 (NRRL B-50131), production propionic acid Propionibacterium P179 (NRRL B-50133), production propionibacterium acide-propionici P169 (PTA 5271), production propionibacterium acide-propionici P170 (PTA 5272), propionibacterium jensenii P63 (NRRL B-30979), propionibacterium jensenii P195 (NRRL B-50132), lactic acid It is galactococcus ID7 (PTA 6103), Lactococcus lactis JD19 (PTA 6104), lactobacillus acidophilus A2020 (NRRL B-30977), thermophilic Lactobacillus lactis A4000h (NRRL B-30978), lactobacillus acidophilus PIBc6 (NRRL B-50103), Lactobacillus brevis LBR 1000 (NRRL B-30982), Lactobacillus casei LC222 (NRRL B-30983), family name's lactobacillus PLCB6 (NRRL B-50518), saliva It is lactobacillus o246i33w (NRRL B-50102), Lactobacillus brevis AJ25 (PTA-6099), Lactobacillus brevis HE17 (PTA-6100), short Lactobacillus 1E-1 (PTA-6509), lactobacillus lactis Cl15 (PTA-6101), lactobacillus lactis DJ6 (PTA-6102), rhamnose Lactobacillus (CNCM I-3698), Lactobacillus farciminis (CNCM I-3699), enterococcus faecium EF141 (EN-1) (NRRL B- 30981), enterococcus faecium 2-1d (NRRL B-50519), Pediococcus acidilactici PIJe3 (NRRL B-50101), Pediococcus acidilactici O246e42 (NRRL B-50171) and combinations thereof.

32. the feed addictive composition of claim 22, wherein phytase are 6-phytase or 3- inositols Six-phosphatase.

33. the feed addictive composition of claim 32, wherein phytase are 6-phytases.

34. the feed addictive composition of claim 22, wherein phytase are escherichia coli phytases, or Buttiauxella phytase or Citrobacter phytase or Hafnia phytase, or Aspergillus phytase or Penicillium phytase or trichoderma phytase or Escherichia coli inositol six Phosphatase or Hansenula phytase or Peniophora category phytase.

35. kit, it includes the feed addictive composition of any one of claim 22 to 34 and operation instructions.

36. the method for preparing feed addictive composition comprising by least one Direct-Fed Microbials and phytic acid Enzyme combines so that the dosage of the composition mysoinositol six-phosphatase：

A. it is 30,000FTU/g compositions or more when being added in feed by least 50g/ tonnes of (MT) feed；

B. it is 20,000FTU/g compositions or more when being added in feed by least 75g/ tonnes of (MT) feed；

C. it is 15,000FTU/g compositions or more when being added in feed by least 100g/ tonnes of (MT) feed；

D. it is 15,000FTU/g compositions or more when being added in feed by least 100g/ tonnes of (MT) feed；

E. it is 10,000FTU/g compositions or more when being added in feed by least 150g/ tonnes of (MT) feed；

F. it is 7,500FTU/g compositions or more when being added in feed by least 200g/ tonnes of (MT) feed；

G. it is 5,000FTU/g compositions or more when being added in feed by least 300g/ tonnes of (MT) feed；

And the dosage of Direct-Fed Microbials described in the feed addictive composition is from 2.5x10³CFU DFM:1FTU enzymes To 6.7x10⁶CFU:In the range of 1FTU enzymes.

37. the method for claim 36, wherein the method further include packaging step.

38. the method for claim 36, the wherein dosage of Direct-Fed Microbials described in feed addictive composition from 3.8x10³CFU DFM:1FTU enzymes are to 2.0x10⁵CFU:In the range of 1FTU enzymes.

39. feed, it includes the feed addictive compositions of any one of claim 22-34.

40. the feed of claim 39, wherein phytase press 1500FTU/kg feeds and 20,000FTU//kg feeds it Between dosage exist.

41. the feed of claim 39 or claim 40, wherein DFM press 7.5x10⁴CFU/kg feeds and 1x10⁷CFU/kg is raised The dosage of material exists.

42. the method for preparing feed comprising combine feed ingredient with the feed addictive of any one of claim 22-34 Object mixes.

43. pre-composition, described it includes feed addictive composition and at least one minerals and/or at least one vitamin Feed addictive composition contains the combination of at least one Direct-Fed Microbials and phytase, wherein phytic acid Enzyme is by existing below in the pre-composition：

And the Direct-Fed Microbials are pressed from 2.5x10³CFU DFM:1FTU enzymes are to 6.7x10⁶CFU:The range of 1FTU enzymes exists In the pre-composition.

44. pre-composition, it includes the feed addictive compositions of any one of claim 22-34 and at least one minerals And/or the combination of at least one vitamin.

45. the feed addictive composition of any one of claim 22-34, the pre-composition of claim 43 or 44 or right are wanted Seek use of the feed of any one of 39-41 in preparing the drug for preventing and/or treating necrotic enteritis and/or globidiosis On the way.