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Title:
COMPOSITIONS AND METHODS OF IMPROVING THE DIGESTIBILITY OF ANIMAL FEED
Document Type and Number:
WIPO Patent Application WO/2016/060935
Kind Code:
A2
Abstract:
The present invention relates to methods and compositions comprising one or more bacteria for improving the in vitro true digestibility and/or neutral detergent fiber digestibility of an animal feed.

Inventors:
LEWIS, Derrick (Novozymes North America, Inc.108 TW Alexander Drive, Building 1, Durham North Carolina, 27709, US)
KOFOD, Lene Venke (Novozymes A/S, Krogshoejvej 36, Bagsvaerd, Bagsvaerd, DK)
HAUGHNEY, Shannon Lee (Novozymes North America, Inc.108 TW Alexander Drive, Building 1, Durham North Carolina, 27709, US)
Application Number:
US2015/054718
Publication Date:
April 21, 2016
Filing Date:
October 08, 2015
Export Citation:
Click for automatic bibliography generation   Help
Assignee:
NOVOZYMES A/S (Krogshoejvej 36, Bagsvaerd, Bagsvaerd, DK)
LEWIS, Derrick (Novozymes North America, Inc.108 TW Alexander Drive, Building 1, Durham North Carolina, 27709, US)
International Classes:
A23K1/00
Domestic Patent References:
2010-03-25
2011-09-22
1998-07-02
2000-07-27
2003-08-14
2003-05-30
2003-06-12
1994-01-20
2002-11-14
2008-02-14
2006-04-06
1997-02-13
1998-12-10
1998-12-10
2000-11-23
2001-01-04
Foreign References:
US5093121A1992-03-03
EP0495725A11992-07-22
US4919936A1990-04-24
Other References:
PENG ET AL., JOURNAL OF ANIMAL PHYSIOLOGY AND ANIMAL NUTRITION, vol. 96, 2011, pages 506 - 512
USING NDF DIGESTIBILITY IN RATION FORMULATION, HOFFMAN AND COMBS, FOCUS ON FORAGE, vol. 6, no. 3
"Nomenclature Committee of the International Union of Biochemistry and Molecular Biology (IUB-MB", 1992, ACADEMIC PRESS, INC.
BAIROCH: "ENZYME database", NUCLEIC ACIDS RES, vol. 28, 2000, pages 304 - 305
COUTINHO, P.M.; HENRISSAT, B., CARBOHYDRATE-ACTIVE ENZYMES SERVER, 1999
COUTINHO, P.M.; HENRISSAT, B.: "Recent Advances in Carbohydrate Bioengineering", 1999, THE ROYAL SOCIETY OF CHEMISTRY, article "Carbohydrate-active enzymes: an integrated database approach", pages: 3 - 12, XP008041544
COUTINHO, P.M.; HENRISSAT, B.: "Genetics, Biochemistry and Ecology of Cellulose Degradation", 1999, UNI PUBLISHERS CO., article "The modular structure of cellulases and other carbohydrate-active enzymes: an integrated database approach", pages: 15 - 23
JOURNAL OF DAIRY SCIENCE, vol. 74, 1991, pages 3583 - 3597
STERN; HOOVER: "Proc. Continuous Culture Fermenters: Frustration or Fermentation", 1990, AM. DAIRY SCI. ASSOC., pages: 17 - 32
WELLER; PILGRIM: "Passage of protozoa and volatile fatty acids from the rumen of sheep and from a continuous in vitro fermentation system", BR. J. NUTR., vol. 32, 1974, pages 341 - 352
VAN SOEST: "Methods for dietary fiber, neutral detergent fiber, and nonstarch polysaccharides in relation to animal nutrition", J. DAIRY SCI., vol. 74, 1991, pages 3583 - 3597
STERN; HOOVER: "Proc. Continuous Culture Fermenters: Frustration or Fermentation", 1990, AM. DAIRY SCI. ASSOC., article "The dual flow continuous culture system", pages: 17 - 32
Attorney, Agent or Firm:
LAMBIRIS, Elias et al. (Novozymes North America, Inc.60 East 42nd Street, Suite 70, New York New York, 10165, US)
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Claims:
Claims

1. A method for improving the digestibility of a ruminant feed, comprising adding to the ruminant feed a Bacillus strain selected from the group consisting of Bacillus pumilus, Bacillus amyloliquefaciens, Bacillus subtilis and Bacillus licheniformis wherein the Bacillus strain improves the in vitro true digestibility and/or neutral detergent fiber digestibility of the ruminant feed and wherein the Bacillus strain has enzyme activity against Xylan under aerobic conditions. 2. A method for improving the digestibility of a ruminant feed, comprising adding to the ruminant feed a Bacillus strain selected from the group consisting of:

the strain having the deposit accession number ATCC 700385 or a strain having all of the identifying characteristics of Bacillus ATCC 700385 or a mutant thereof;

the strain having the deposit accession number NRRL B-50136 or a strain having all of the identifying characteristics of Bacillus NRRL B-50136 or a mutant thereof;

the strain having the deposit accession number NRRL B-50147 or a strain having all of the identifying characteristics of Bacillus NRRL B-50147 or a mutant thereof;

the strain having the deposit accession number NRRL B-50622 or a strain having all of the identifying characteristics of Bacillus NRRL B-50622 or a mutant thereof;

the strain having the deposit accession number NRRL B-50623 or a strain having all of the identifying characteristics of Bacillus NRRL B-50623 or a mutant thereof;

the strain having the deposit accession number NRRL B-50605 or a strain having all of the identifying characteristics of Bacillus NRRL B-50605 or a mutant thereof;

the strain having the deposit accession number NRRL B-50621 or a strain having all of the identifying characteristics of Bacillus NRRL B-50621 or a mutant thereof;

the strain having the deposit accession number NRRL B-50015 or a strain having all of the identifying characteristics of Bacillus NRRL B-50015 or a mutant thereof;

the strain having the deposit accession number NRRL B-50016 or a strain having all of the identifying characteristics of Bacillus NRRL B-50016 or a mutant thereof;

the strain having the deposit accession number NRRL B-50885 or a strain having all of the identifying characteristics of Bacillus NRRL B-50885 or a mutant thereof;

the strain having the deposit accession number NRRL B-50886 or a strain having all of the identifying characteristics of Bacillus NRRL B-50886 or a mutant thereof;

the strain having the deposit accession number NRRL B-50013 or a strain having all of the identifying characteristics of Bacillus NRRL B-50013 or a mutant thereof;

the strain having the deposit accession number NRRL B-50141 or a strain having all of the identifying characteristics of Bacillus NRRL B-50141 or a mutant thereof; the strain having the deposit accession number NRRL B-50151 or a strain having all of the identifying characteristics of Bacillus NRRL B-50151 or a mutant thereof;

the strain having the deposit accession number NRRL B-50606 or a strain having all of the identifying characteristics of Bacillus NRRL B-50606 or a mutant thereof;

the strain having the deposit accession number PTA-7543 or a strain having all of the identifying characteristics of Bacillus PTA-7543 or a mutant thereof;

the strain having the deposit accession number PTA-7547 or a strain having all of the identifying characteristics of Bacillus PTA-7547 or a mutant thereof;

the strain having the deposit accession number NRRL B-50888 or a strain having all of the identifying characteristics of Bacillus NRRL B-50888 or a mutant thereof;

the strain having the deposit accession number PTA-7549 or a strain having all of the identifying characteristics of Bacillus PTA-7549 or a mutant thereof;

the strain having the deposit accession number NRRL B-50349 or a strain having all of the identifying characteristics of Bacillus NRRL B-50349 or a mutant thereof;

the strain having the deposit accession number PTA-7541 or a strain having all of the identifying characteristics of Bacillus PTA-7541 or a mutant thereof;

the strain having the deposit accession number NRRL B-59648 or a strain having all of the identifying characteristics of Bacillus NRRL B-59648 or a mutant thereof;

the strain having the deposit accession number NRRL B-59650 or a strain having all of the identifying characteristics of Bacillus NRRL B-59650 or a mutant thereof;

the strain having the deposit accession number NRRL B-59651 or a strain having all of the identifying characteristics of Bacillus NRRL B-59651 or a mutant thereof;

the strain having the deposit accession number NRRL B-676141 or a strain having all of the identifying characteristics of Bacillus NRRL B-676141 or a mutant thereof; and

the strain having the deposit accession number NRRL B-676140 or a strain having all of the identifying characteristics of Bacillus NRRL B-676140 or a mutant thereof; or

any combination thereof.

3. A method for improving the digestibility of a ruminant feed, comprising adding to the ruminant feed a Bacillus strain selected from the group consisting of:

the strain having the deposit accession number NRRL B-50605 or a strain having all of the identifying characteristics of Bacillus NRRL B-50605 or a mutant thereof;

the strain having the deposit accession number NRRL B-50885 or a strain having all of the identifying characteristics of Bacillus NRRL B-50885 or a mutant thereof;

the strain having the deposit accession number NRRL B-50886 or a strain having all of the identifying characteristics of Bacillus NRRL B-50886 or a mutant thereof; the strain having the deposit accession number PTA-7541 or a strain having all of the identifying characteristics of Bacillus PTA-7541 or a mutant thereof;

the strain having the deposit accession number NRRL B-59648 or a strain having all of the identifying characteristics of Bacillus NRRL B-59648 or a mutant thereof;

the strain having the deposit accession number NRRL B-59650 or a strain having all of the identifying characteristics of Bacillus NRRL B-59650 or a mutant thereof;

the strain having the deposit accession number NRRL B-59651 or a strain having all of the identifying characteristics of Bacillus NRRL B-59651 or a mutant thereof;

the strain having the deposit accession number NRRL B-676141 or a strain having all of the identifying characteristics of Bacillus NRRL B-676141 or a mutant thereof; and

the strain having the deposit accession number NRRL B-676140 or a strain having all of the identifying characteristics of Bacillus NRRL B-676140 or a mutant thereof; or any combination thereof. 4. A method for improving the digestibility of a ruminant feed comprising:

(a) feeding a ruminant; and

(b) administering to a ruminant a Bacillus strain selected from the group consisting of Bacillus pumilus, Bacillus amyloliquefaciens, Bacillus subtilis and Bacillus licheniformis wherein the Bacillus strain improves the in vitro true digestibility and/or neutral detergent fiber digestibility of the animal feed and wherein the Bacillus strain has enzyme activity against Xylan under aerobic conditions;

and wherein step (a) occurs before, after, or simultaneously with step (b).

5. A method for improving the digestibility of a ruminant feed comprising:

(a) feeding a ruminant; and

(b) administering to a ruminant a Bacillus strain selected from the group consisting of:

the strain having the deposit accession number ATCC 700385; or a strain having all of the identifying characteristics of Bacillus ATCC 700385 or a mutant thereof;

the strain having the deposit accession number NRRL B-50136; or a strain having all of the identifying characteristics of Bacillus NRRL B-50136 or a mutant thereof;

the strain having the deposit accession number NRRL B-50147; or a strain having all of the identifying characteristics of Bacillus NRRL B-50147 or a mutant thereof; the strain having the deposit accession number NRRL B-50622; or a strain having all of the identifying characteristics of Bacillus NRRL B-50622 or a mutant thereof;

the strain having the deposit accession number NRRL B-50623; or a strain having all of the identifying characteristics of Bacillus NRRL B-50623 or a mutant thereof;

the strain having the deposit accession number NRRL B-50605; or a strain having all of the identifying characteristics of Bacillus NRRL B-50605 or a mutant thereof;

the strain having the deposit accession number NRRL B-50621 ; or a strain having all of the identifying characteristics of Bacillus NRRL B-50621 or a mutant thereof;

the strain having the deposit accession number NRRL B-50015; or a strain having all of the identifying characteristics of Bacillus NRRL B-50015 or a mutant thereof;

the strain having the deposit accession number NRRL B-50016; or a strain having all of the identifying characteristics of Bacillus NRRL B-50016 or a mutant thereof;

the strain having the deposit accession number NRRL B-50885; or a strain having all of the identifying characteristics of Bacillus NRRL B-50885 or a mutant thereof;

the strain having the deposit accession number NRRL B-50886; or a strain having all of the identifying characteristics of Bacillus NRRL B-50886 or a mutant thereof;

the strain having the deposit accession number NRRL B-50013; or a strain having all of the identifying characteristics of Bacillus NRRL B-50013 or a mutant thereof;

the strain having the deposit accession number NRRL B-50141 ; or a strain having all of the identifying characteristics of Bacillus NRRL B-50141 or a mutant thereof;

the strain having the deposit accession number NRRL B-50151 ; or a strain having all of the identifying characteristics of Bacillus NRRL B-50151 or a mutant thereof;

the strain having the deposit accession number NRRL B-50606; or a strain having all of the identifying characteristics of Bacillus NRRL B-50606 or a mutant thereof; the strain having the deposit accession number PTA-7543; or a strain having all of the identifying characteristics of Bacillus PTA-7543 or a mutant thereof;

the strain having the deposit accession number PTA-7547; or a strain having all of the identifying characteristics of Bacillus PTA-7547 or a mutant thereof;

the strain having the deposit accession number NRRL B-50888; or a strain having all of the identifying characteristics of Bacillus NRRL B-50888 or a mutant thereof;

the strain having the deposit accession number PTA-7549; or a strain having all of the identifying characteristics of Bacillus PTA-7549 or a mutant thereof;

the strain having the deposit accession number NRRL B-50349; or a strain having all of the identifying characteristics of Bacillus NRRL B-50349 or a mutant thereof,

the strain having the deposit accession number PTA-7541 or a strain having all of the identifying characteristics of Bacillus PTA-7541 or a mutant thereof;

the strain having the deposit accession number NRRL B-59648 or a strain having all of the identifying characteristics of Bacillus NRRL B-59648 or a mutant thereof;

the strain having the deposit accession number NRRL B-59650 or a strain having all of the identifying characteristics of Bacillus NRRL B-59650 or a mutant thereof;

the strain having the deposit accession number NRRL B-59651 or a strain having all of the identifying characteristics of Bacillus NRRL B-59651 or a mutant thereof;

the strain having the deposit accession number NRRL B-676141 or a strain having all of the identifying characteristics of Bacillus NRRL B-676141 or a mutant thereof; and

the strain having the deposit accession number NRRL B-676140 or a strain having all of the identifying characteristics of Bacillus NRRL B-676140 or a mutant thereof; or any combination thereof, and

wherein step (a) occurs before, after, or simultaneously with step (b).

6. The method of any of claims 1 to 5, wherein the ruminant feed has a 48 hour in vitro NDF Digestibility of 30-70% of NDF such as a 48 hour in vitro NDF Digestibility of NDF selected from the group consisting of from 30-40%, for example from 40-50%, such as from 50-60%, for example from 60-70%, or any combination of these intervals.

7. The method of any of claims 1 to 6, wherein the ruminant feed comprises forage.

8. The method of any of claims 1 to 7, wherein the ruminant feed comprises roughage.

9. The method of any of claims 1 to 8, wherein the method increases the milk yield and/or the meat yield of the ruminant.

10. The method of any of claims 1 to 9, wherein the bacterial count of each of the bacterial strains is between 1x104 and 1 x1014 CFU/kg of forage, preferably between 1 x106 and 1 x1012 CFU/kg of forage, and more preferably between 1 x107 and 1 x1011 CFU/kg of forage and/or between 1x105 and 1 x1015 CFU/animal/day, preferably between 1 x107 and 1x1013 CFU/animal/day, and more preferably between 1x108 and 1x1012 CFU/animal/day.

1 1 . An animal feed composition such as a ruminant feed composition comprising forage and a Bacillus strain selected from the group consisting of:

the strain having the deposit accession number NRRL B-50885; or a strain having all of the identifying characteristics of NRRL B-50885 or a mutant thereof;

the strain having the deposit accession number NRRL B-50886; or a strain having all of the identifying characteristics of NRRL B-50886 or a mutant thereof;

the strain having the deposit accession number NRRL B-50888 or a strain having all of the identifying characteristics of NRRL B-50888 or a mutant thereof;

the strain having the deposit accession number PTA-7541 or a strain having all of the identifying characteristics of Bacillus PTA-7541 or a mutant thereof;

the strain having the deposit accession number NRRL B-59648 or a strain having all of the identifying characteristics of Bacillus NRRL B-59648 or a mutant thereof;

the strain having the deposit accession number NRRL B-59650 or a strain having all of the identifying characteristics of Bacillus NRRL B-59650 or a mutant thereof;

the strain having the deposit accession number NRRL B-59651 or a strain having all of the identifying characteristics of Bacillus NRRL B-59651 or a mutant thereof;

the strain having the deposit accession number NRRL B-676141 or a strain having all of the identifying characteristics of Bacillus NRRL B-676141 or a mutant thereof; and

the strain having the deposit accession number NRRL B-676140 or a strain having all of the identifying characteristics of Bacillus NRRL B-676140 or a mutant thereof; or any combination thereof. 12. The animal feed composition of claim 1 1 , wherein one or more of the bacterial strains are in spore form.

13. The animal feed composition of claim 1 1 or 12, wherein the bacterial count of each of the bacterial strains is between 1 x104 and 1 x1014 CFU/kg of forage, preferably between 1x106 and 1 x1012 CFU/kg of forage, and more preferably between 1x107 and 1x1011 CFU/kg of forage.

14. The animal feed composition of any of claims 1 1 to 13 further comprising roughage.

15. A Bacillus strain selected from the group consisting of

the strain having the deposit accession number PTA-7541 or a strain having all of the identifying characteristics of Bacillus PTA-7541 or a mutant thereof;

the strain having the deposit accession number NRRL B-676141 or a strain having all of the identifying characteristics of Bacillus NRRL B-676141 or a mutant thereof; and

the strain having the deposit accession number NRRL B-676140 or a strain having all of the identifying characteristics of Bacillus NRRL B-676140 or a mutant thereof.

Description:
COMPOSITIONS AND METHODS OF IMPROVING

THE DIGESTIBILITY OF ANIMAL FEED

Reference to a Deposit of Biological Material

This application contains a reference to a deposit of biological material, which deposit is incorporated herein by reference.

BACKGROUND OF THE INVENTION

Field of the Invention

The present invention relates to methods and compositions comprising one or more bacteria for improving the in vitro true digestibility and/or neutral detergent fiber digestibility of an animal feed.

Background of the Invention

Feed ration costs account for 45-60 percent of the total cost of producing milk, so optimal nutrition is important. Ideally, appropriate nutrient levels should be maintained, while feed costs are carefully maintained. Such optimal nutrition will enhance milk production, improve overall health of the ruminant, and reduce associated costs like veterinary bills, drug treatments, and breeding.

The main nutrient categories of importance for ruminants are carbohydrates, fats, proteins, minerals, vitamins, and water. Through a process of rumination, the ruminant reduces the particle size of feed in the rumen, which enhances microbial function, and allows for easier passage out of the stomach compartments. In ruminants a major part of carbohydrates are derived from plant fibers which are degraded in the rumen by the rumen microflora. The carbohydrates are then fermented to acids which serve as nutrients for the ruminant. The degraded and fermented content of the rumen then passes to the reticulum and the rest of the ruminant's digestive tract for further degradation and absorption of the released nutrients.

The neutral detergent fiber ("NDF") from forage is thus an important component in many ruminant diets and a major source of energy for the ruminant. Forage NDF also stimulates chewing and secretion of salivary buffers to neutralize the fermentation acids in the rumen. However, the NDF is typically only partly degraded and fermented in the rumen and the un-degraded NDF pass through the ruminant and the energy contained within the excreta is therefore lost.

It is thus an object of the invention to provide solutions which improve the digestibility of neutral detergent fiber whilst maintaining an optimal function of the rumen for fermentation of the digested NDF. Description of the Related Art

US 5,093,121 describes the use of lactic acid producing bacteria to increase the protein content in cow milk.

EP 0495725 describes the process of increasing the microbial population in the rumen by feeding the ruminant with a Bacillus cereus spores, thereby improving milk and meat production in the ruminant.

WO 2010/033714 describes a method for enhancing the health of an animal comprising administering to the animal a composition comprising Bacillus subtilis QST713.

Peng et al. {Journal of Animal Physiology and Animal Nutrition 96: 506-512 (201 1 )) describe the effect of feeding Bacillus subtilis natto fermentation product on milk production and composition, blood metabolites and rumen fermentation in early lactation dairy cows.

WO 201 1/1 15306 describes the use of microbes from Bacillus subtilis to improve the feed utilisation efficiency in ruminants.

US 4,919,936 describes a method for increasing the weight gain in animals comprising feeding an animal a probiotic comprising Bacillus subtilis C-3102.

Summary of the Invention

It has been surprisingly found that the addition of direct fed microbes (DFM) from Bacillus species to animal feed improves the in vitro true digestibility (IVTD) and/or the neutral detergent fiber digestibility (NDFD) and/or the Acid Detergent Fiber Digestibility (ADFD) of the feed compared to feed without any DFM added.

The DFM of the present invention has protease activity. The protease activity of DFM improves delignification of e.g. corn stover by acting on protein network. Ultimately, the protease activity of the DFM provides better accessibility for the cellulose and xyaln components increasing digestibility of the stover material when used as an animal feed such as a ruminant feed.

In one embodiment the invention relates to a method for improving the digestibility of an animal feed, comprising adding to the animal feed a Bacillus strain selected from the group consisting of Bacillus pumilus, Bacillus amyloliquefaciens, Bacillus subtilis and Bacillus licheniformis wherein the Bacillus strain improves the in vitro true digestibility and/or neutral detergent fiber digestibility and/or the Acid Detergent Fiber Digestibility (ADFD) of the animal feed and wherein the Bacillus strain has enzyme activity against Casein and/or Xylan under aerobic conditions.

The invention also relates to a method for improving the digestibility of an animal feed, comprising adding to the animal feed one or more Bacillus strains selected from the group consisting of the strain having the deposit accession number ATCC 700385, NRRL B- 50136, NRRL B-50147, NRRL B-50622, NRRL B-50623, NRRL B-50605, NRRL B-50621 , NRRL B-50015, NRRL B-50016, NRRL B-50885, NRRL B-50886, NRRL B-50013, NRRL B- 50141 , NRRL B-50151 , NRRL B-50606, PTA-7543, PTA-7547, NRRL B-50888, PTA-7549 NRRL B-50349, PTA-7541 , NRRL B-59648, NRRL B-59650, NRRL B-59651 , NRRL B- 676141 , and NRRL B-676140 or any combination thereof, wherein the Bacillus strain improves the in vitro true digestibility and/or neutral detergent fiber digestibility and/or the Acid Detergent Fiber Digestibility (ADFD) of the animal feed. In an embodiment, the invention relates to an animal feed that is fed to a ruminant; comprises forage and optionally other components; improves the digestibility of the animal feed; and increases milk and/or meat yield.

The invention also relates to a method for improving the digestibility of an animal feed such as a ruminant feed comprising:

(a) feeding an animal a feed such as a ruminant feed; and

(b) administering to an animal such as a ruminant a Bacillus strain selected from the group consisting of Bacillus pumilus, Bacillus amyloliquefaciens, Bacillus subtilis and

Bacillus licheniformis, or any combination thereof, wherein the Bacillus strain improves the in vitro true digestibility and/or neutral detergent fiber digestibility and/or the Acid Detergent Fiber Digestibility (ADFD) of the animal feed and wherein the Bacillus strain has enzyme activity against Casein and/or Xylan under aerobic conditions.

In a further aspect, the invention relates to a method for improving the digestibility of an animal feed such as a ruminant feed comprising:

(a) feeding an animal a feed such as a ruminant feed; and

(b) administering to an animal such as a ruminant one or more Bacillus strains selected from the group consisting of the strain having the deposit accession number ATCC 700385, NRRL B-50136, NRRL B-50147, NRRL B-50622, NRRL B-50623, NRRL B-50605, NRRL B-50621 , NRRL B-50015, NRRL B-50016, NRRL B-50885, NRRL B-50886, NRRL B- 50013, NRRL B-50141 , NRRL B-50151 , NRRL B-50606, PTA-7543, PTA-7547, NRRL B- 50888, PTA-7549 NRRL B-50349, PTA-7541 , NRRL B-59648, NRRL B-59650, NRRL B- 59651 , NRRL B-676141 , and NRRL B-676140 or any combination thereof, wherein the Bacillus strain improves the in vitro true digestibility and/or neutral detergent fiber digestibility and/or the Acid Detergent Fiber Digestibility (ADFD) of the animal feed, and wherein step (a) occurs before, after, or simultaneously with step (b). In an embodiment, the method relates to an animal feed that is fed to a ruminant; and comprises forage and optionally other components.

In a further aspect, the invention relates to an animal feed composition such as a ruminant feed comprising forage and one or more bacteria culture, wherein the bacteria culture is selected from the group consisting of the strains having the deposit accession number NRRL B-50885, NRRL B-50886 NRRL B-50888, PTA-7541 , NRRL B-59648, NRRL B-59650, NRRL B-59651 , NRRL B-676141 , and NRRL B-676140 or any combination thereof. In an embodiment, the animal feed composition comprises one of more additional bacterium; is fed to a ruminant; and further comprises concentrate, one or more enzymes, one or more additional microbes, one or more vitamins, one or more minerals, one or more amino acids, and one or more other feed ingredients.

Definitions

In general, the terms and phrases used herein have their art-recognized meaning, which can be found by reference to standard texts, journal references, and context known to those skilled in the art. The following definitions are provided to clarify their specific use in context of the disclosure.

As used herein, the singular forms "a", "an" and "the" are intended to include the plural forms as well, unless the context clearly indicates otherwise.

Acid Detergent Fiber (ADF): The term "acid detergent fiber" is a measure of the portion of the diet that is comprised of cellulose and lignin.

Acid Detergent Fiber Digestibility (ADFD): The term "acid detergent fiber digestibility" means an anaerobic fermentation performed in the laboratory to simulate digestion as it occurs in the rumen. Rumen fluid is collected from ruminally cannulated animals, such as high producing dairy cows consuming a typical total mixed ration (TMR). Forage samples are incubated in rumen fluid and buffer for a specified time period at 39°C (body temperature). During this time, the microbial population in the rumen fluid digests the sample as would occur in the rumen. Upon completion, the samples are extracted in acidic detergent to leave behind the undigested fibrous residue. The result is a measure of digestibility that can be used to estimate energy. ADFD can be determined using the acid detergent fiber digestibility assay as described in the examples.

Animal feedB: The term "animal feed" refers to any compound, preparation, or mixture suitable for, or intended for intake by an animal. Animal feed for ruminants generally comprises forage (including roughage and silage) and may further comprise concentrates as well as vitamins, minerals, enzymes and/or other feed ingredients (such as in a premix).

Blend: the term "blend" means more than one of the bacterial strains described herein.

Composition: The term "composition" refers to a composition comprising a carrier and at least one bacterial strain as described herein. The compositions described herein may be mixed with an animal feed(s) and referred to as a "mash feed."

Concentrates: The term "concentrates" means feed with high protein and energy concentrations, such as fish meal, molasses, oligosaccharides, sorghum, seeds and grains (either whole or prepared by crushing, milling, etc from e.g. corn, oats, rye, barley, wheat), oilseed press cake (e.g. from cottonseed, safflower, sunflower, soybean, rapeseed/canola, peanut or groundnut), palm kernel cake, yeast derived material and distillers grains (such as wet distillers grains (WDS) and dried distillers grains with solubles (DDGS)).

Direct Fed Microbial: The term "direct fed microbial" means live micro-organisms including spores which, when administered in adequate amounts, confer a benefit, such as improved digestion or health, on the host.

Effective amount/concentration/dosage: The terms "effective amount", "effective concentration", or "effective dosage" are defined as the amount, concentration, or dosage of the bacterial strain(s) sufficient to improve the digestion or yield of an animal. The actual effective dosage in absolute numbers depends on factors including: the state of health of the animal in question, other ingredients present. The "effective amount", "effective concentration", or "effective dosage" of the bacterial strains may be determined by routine assays known to those skilled in the art.

Forage: The term "forage" as defined herein also includes roughage. Forage is fresh plant material such as hay and silage from forage plants, grass and other forage plants, seaweed, sprouted grains and legumes, or any combination thereof. Examples of forage plants are Alfalfa (lucerne), birdsfoot trefoil, brassica (e.g. kale, rapeseed (canola), rutabaga (swede), turnip), clover (e.g. alsike clover, red clover, subterranean clover, white clover), grass (e.g. Bermuda grass, brome, false oat grass, fescue, heath grass, meadow grasses, orchard grass, ryegrass, Timothy-grass), corn (maize), millet, barley, oats, rye, sorghum, soybeans and wheat and vegetables such as beets. Forage further includes crop residues from grain production (such as corn stover; straw from wheat, barley, oat, rye and other grains); residues from vegetables like beet tops; residues from oilseed production like stems and leaves form soy beans, rapeseed and other legumes; and fractions from the refining of grains for animal or human consumption or from fuel production or other industries.

Improves the digestibility of the animal feed: The term "improves the digestibility of the animal feed" means a method and/or composition that improves the digestibility of the animal feed by increasing the breakdown of the hemicellulose and/or cellulose of the animal feed in the digestive tract/rumen in order to provide more energy to the animal.

In vitro True Digestibility (IVTD): The term "in vitro true digestibility" means an anaerobic fermentation performed in the laboratory to simulate digestion as it occurs in the rumen. Rumen fluid is collected from ruminally cannulated animals, such as high producing dairy cows consuming a typical total mixed ration (TMR). Forage samples are incubated in rumen fluid and buffer for a specified time period at 39°C (body temperature). During this time, the microbial population in the rumen fluid digests the sample as would occur in the rumen. Upon completion, the samples are extracted in neutral detergent solution to leave behind the undigested fibrous residue. The result is a measure of digestibility that can be used to estimate energy. IVTD can be determined using the in vitro true digestibility assay as described in the examples.

Isolated: The term "isolated" means that the one or more bacterial strains described herein are in a form or environment which does not occur in nature, that is, the one or more bacterial strains are at least partially removed from one or more or all of the naturally occurring constituents with which it is associated in nature.

Neutral Detergent Fiber (NDF): The term "neutral detergent fiber" is a measure of hemicellulose, cellulose and lignin representing the fibrous bulk of the forage. These three components are classified as cell wall or structural carbohydrates. They give the plant rigidity enabling it to support itself as it grows, much like the skeleton in animals. Hemicellulose and cellulose can be broken down by microbes in the rumen to provide energy to the animal.

Neutral Detergent Fiber Digestibility (NDFD): The term "neutral detergent fiber digestibility" the proportion of NDF potentially available as determined by an in vitro incubation. NDFD is expressed as a percentage of the NDF. The NDFD can be used to rank forages on potential fiber digestibility and in energy calculations. NDFD can be determined using the neutral detergent fiber digestibility assay as described in the examples.

Pellet: The terms "pellet" and/or "pelleting" refer to solid rounded, spherical and/or cylindrical tablets or pellets and the processes for forming such solid shapes, particularly feed pellets and solid extruded animal feed. As used herein, the terms "extrusion" or

"extruding" are terms well known in the art and refer to a process of forcing a composition, as described herein, through an orifice under pressure.

pH: The term "pH" is a measure of the degree of acidity. Good corn silage typically has a pH of 3.5 - 4.5 and hay crop silages 3.8 - 5.3.

Roughage: The term "roughage" means dry plant material with high levels of fiber, such as fiber, bran, husks from seeds and grains and crop residues (such as stover, copra, straw, chaff, sugar beet waste).

Ruminant: The term "ruminant" means a mammal that digests plant-based food by initially fermenting/degrading it within the animal's first compartment of the stomach, principally through bacterial actions, then regurgitating the semi-digested mass, now known as cud, and chewing it again. The process of re-chewing the cud to further break down plant matter and stimulate digestion is called "ruminating". Examples of ruminants are cattle, cow, beef cattle, young calf, goat, sheep, lamb, deer, yank, camel and llama. Preferred examples are cattle, cow, beef cattle and young calf.

Silage: The term "silage" means fermented, high-moisture stored fodder which can be fed to ruminants (cud-chewing animals such as cattle and sheep) or used as a biofuel feedstock for anaerobic digesters. It is fermented and stored in a process called ensilage, ensiling or silaging, and is usually made from grass or cereal crops (e.g. maize, sorghum, oats, rye, timothy etc forage grass plants),) or legume crops like clovers/trefoils, alfalfa, vetches, using the entire green plant (not just the grain). Silage can be made from many field crops, and special terms may be used depending on type (oatlage for oats, haylage for alfalfa). Silage is made either by placing cut green vegetation in a silo, by piling it in a large heap covered with plastic sheet, or by wrapping large bales in plastic film.

Spore: The terms "spore" and "endospore" are interchangeable and have their normal meaning which is well known and understood by those of skill in the art. As used herein, the term spore refers to a microorganism in its dormant, protected state.

Stable: The term "stable" is a term that is known in the art, and in a preferred aspect, stable is intended to mean the ability of the microorganism to remain in a spore form until it is administered to an animal to improve the health of the animal.

Sub-optimal animal feed: The terms "sub-optimal animal feed" and "animal feed is sub-optimal" means an animal feed that contains a high level of un-digestible organic matter, such as the animal feed comprises at least 30%, preferably at least 40%, more preferably at least 50%, even more preferably at least 60% and most preferably at least 70% un-digestible organic matter. Organic Matter (OM) digestibility for ruminants of different types of animal feed can be obtained by looking at reference texts such as e.g. www.feedipedia.org.

Vegetable protein: The term "vegetable protein" refers to any compound, preparation or mixture that includes at least one protein derived from or originating from a vegetable, including modified proteins and protein-derivatives.

Figure legends

Figure 1 shows IVTD and NDFD results obtained by the "In vitro True Digestibility and Neutral Detergent Fiber Digestibility" assay described in Example 1 and presented in Table 1.

Figure 2 shows IVTD and NDFD results obtained by the "In vitro True Digestibility and Neutral Detergent Fiber Digestibility" assay described in Example 1 and presented in Table 2.

Figure 3 shows IVTD and NDFD results obtained by the "In vitro True Digestibility and

Neutral Detergent Fiber Digestibility" assay described in Example 1 and presented in Table 3.

Figure 4 shows IVTD and NDFD results obtained by the "In vitro True Digestibility and Neutral Detergent Fiber Digestibility" assay described in Example 2 and presented in Table 4. Figure 5 shows IVTD and NDFD results obtained by the "In vitro True Digestibility and Neutral Detergent Fiber Digestibility" assay described in Example 2 and presented in Table 5.

Figure 6 shows IVTD and NDFD results obtained by the "In vitro True Digestibility and Neutral Detergent Fiber Digestibility" assay described in Example 2 and presented in Table

6.

Detailed Description of the Invention

It has been surprisingly found that the addition of direct fed microbial (DFM) from Bacillus species to animal feed such as a ruminant feed improves the in vitro true digestibility (IVTD) and/or the neutral detergent fiber digestibility (NDFD) and/or the Acid Detergent Fiber Digestibility (ADFD) of the feed compared to feed in which DFM has not been added.

Methods for improving the in vitro true digestibility and/or neutral detergent fiber digestibility of an animal feed and/or the Acid Detergent Fiber Digestibility (ADFD)

The invention relates to a method for improving the digestibility of an animal feed, comprising adding to the animal feed such as a ruminant feed a Bacillus strain selected from the group consisting of Bacillus pumilus, Bacillus amyloliquefaciens, Bacillus subtilis and Bacillus licheniformis wherein the Bacillus strain improves the in vitro true digestibility and/or neutral detergent fiber digestibility and/or the Acid Detergent Fiber Digestibility (ADFD) of the animal feed and wherein the Bacillus strain has enzyme activity against Casein and/or Xylan under aerobic conditions.

Thus the present invention relates in one embodiment to a method for improving the digestibility of an animal feed, comprising adding to the animal feed one or more Bacillus strains selected from the group consisting of:

the strain having the deposit accession number ATCC 700385;

the strain having the deposit accession number NRRL B- -50136

the strain having the deposit accession number NRRL B- -50147

the strain having the deposit accession number NRRL B- -50622

the strain having the deposit accession number NRRL B- -50623

the strain having the deposit accession number NRRL B- -50605

the strain having the deposit accession number NRRL B- -50621

the strain having the deposit accession number NRRL B- -50015

the strain having the deposit accession number NRRL B- -50016

the strain having the deposit accession number NRRL B- -50885

the strain having the deposit accession number NRRL B- -50886

the strain having the deposit accession number NRRL B- -50013 the strain having the deposit accession number NRRL B-50141 ;

the strain having the deposit accession number NRRL B-50151 ;

the strain having the deposit accession number NRRL B-50606;

the strain having the deposit accession number PTA-7543;

the strain having the deposit accession number PTA-7547;

the strain having the deposit accession number NRRL B-50888;

the strain having the deposit accession number PTA-7549;

the strain having the deposit accession number NRRL B-50349,

the strain having the deposit accession number PTA-7541 ,

the strain having the deposit accession number NRRL B-59648,

the strain having the deposit accession number NRRL B-59650,

the strain having the deposit accession number NRRL B-59651 ,

the strain having the deposit accession number NRRL B-676141 , and

the strain having the deposit accession number NRRL B-676140 or any combination thereof,

wherein the Bacillus strain improves the vitro true digestibility and/or neutral detergent fiber digestibility and/or the Acid Detergent Fiber Digestibility (ADFD) of the animal feed.

In one embodiment the Bacillus strain has enzyme activity under aerobic conditions against one or more substrates selected from the group consisting of amylose, arabinan, arabinoxylan, casein, cellulose and xylan.

In one embodiment the Bacillus strain is a Bacillus amyloliquefaciens strain and has enzyme activity under aerobic conditions against one or more substrates selected from the group consisting of amylose, arabinan, arabinoxylan, casein and xylan.

In one embodiment the Bacillus strain is a Bacillus pumilus strain and has enzyme activity under aerobic conditions against one or more substrates selected from the group consisting of casein and xylan.

In one embodiment the Bacillus strain is a Bacillus subtilis strain and has enzyme activity under aerobic conditions against one or more substrates selected from the group consisting of amylose, arabinan, arabinoxylan, casein and xylan.

In one embodiment the Bacillus strain is a Bacillus licheniformis strain and has enzyme activity under aerobic conditions against one or more substrates selected from the group consisting of amylose, arabinan and casein.

In one embodiment the Bacillus strain has enzyme activity under anaerobic conditions against one or more substrates selected from the group consisting of amylose, arabinan, arabinoxylan, casein, cellulose and xylan. In one embodiment the Bacillus strain is a Bacillus amyloliquefaciens strain and has enzyme activity under anaerobic conditions against one or more substrates selected from the group consisting of amylose, arabinoxylan, casein and xylan.

In one embodiment the Bacillus strain is a Bacillus subtilis strain and has enzyme activity under anaerobic conditions against one or more substrates selected from the group consisting of amylose, arabinan, arabinoxylan and xylan.

In one embodiment the Bacillus strain is a Bacillus licheniformis strain and has enzyme activity under anaerobic conditions against arabinan.

In one embodiment, the invention relates to a method for improving the in vitro true digestibility of an animal feed. In one embodiment the animal is not a human being. In a further embodiment, the animal feed is fed to a ruminant. In a further embodiment, the ruminant is a cattle, cow, beef cattle, young calf, goat, sheep, lamb, deer, yak, camel or llama. In another embodiment, the animal feed further comprises one or more components selected from the list consisting of forage; concentrate; one or more enzymes; one or more additional microbes; one or more vitamins; one or more minerals; one or more amino acids; and one or more other feed ingredients.

In another embodiment, the invention relates to a method for improving the neutral detergent fiber digestibility of an animal feed. In one embodiment the animal is not a human being. In a further embodiment, the animal feed is fed to a ruminant. In a further embodiment, the ruminant is a cattle, cow, beef cattle, young calf, goat, sheep, lamb, deer, yank, camel or llama. In another embodiment, the animal feed further comprises one or more components selected from the list consisting of forage; concentrate; one or more enzymes; one or more additional microbes; one or more vitamins; one or more minerals; one or more amino acids; and one or more other feed ingredients.

In another embodiment, the invention relates to a method for improving the Acid

Detergent Fiber Digestibility (ADFD) of an animal feed. In one embodiment the animal is not a human being. In a further embodiment, the animal feed is fed to a ruminant. In a further embodiment, the ruminant is a cattle, cow, beef cattle, young calf, goat, sheep, lamb, deer, yank, camel or llama. In another embodiment, the animal feed further comprises one or more components selected from the list consisting of forage; concentrate; one or more enzymes; one or more additional microbes; one or more vitamins; one or more minerals; one or more amino acids; and one or more other feed ingredients. In a further embodiment, the method comprises adding to the animal feed one or more bacterial strains such as at least two of the above strains, at least three of the above strains, at least four of the above strains, at least five of the above strains, at least six of the above strains, at least seven of the above strains, at least eight of the above strains, at least nine of the above strains, at least ten of the above strains, at least eleven of the above strains, at least twelve of the above strains, at least thirteen of the above strains, at least fourteen of the above strains, at least fifteen of the above strains, at least sixteen of the above strains, at least seventeen of the above strains, at least eighteen of the above strains, at least nineteen of the above strains, up to and including all of the above strains.

In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers ATCC 700385 and NRRL B-50136. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers ATCC 700385 and NRRL B-50147. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers ATCC 700385 and NRRL B-50622. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers ATCC 700385 and NRRL B-50623. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers ATCC 700385 and NRRL B-50605. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers ATCC 700385 and NRRL B- 50621. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers ATCC 700385 and NRRL B-50015. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers ATCC 700385 and NRRL B-50016. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers ATCC 700385 and NRRL B-50885. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers ATCC 700385 and NRRL B-50886. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers ATCC 700385 and NRRL B-50013. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers ATCC 700385 and NRRL B- 50141 . In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers ATCC 700385 and NRRL B-50151 . In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers ATCC 700385 and NRRL B-50606. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers ATCC 700385 and PTA-7543. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers ATCC 700385 and PTA-7547. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers ATCC 700385 and NRRL B-50888. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers ATCC 700385 and PTA-7549. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers ATCC 700385 and NRRL B-50349.

In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50136 and NRRL B-50147. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50136 and NRRL B-50622. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50136 and NRRL B-50623. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50136 and NRRL B-50605. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50136 and NRRL B-50621 . In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50136 and NRRL B-50015. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50136 and NRRL B-50016. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50136 and NRRL B-50885. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50136 and NRRL B-50886. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50136 and NRRL B-50013. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50136 and NRRL B-50141 . In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50136 and NRRL B-50151 . In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50136 and NRRL B-50606. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50136 and PTA-7543. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50136 and PTA-7547. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50136 and NRRL B-50888. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50136 and PTA-7549. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50136 and NRRL B- 50349. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50147 and NRRL B-50622. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50147 and NRRL B-50623. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50147 and NRRL B-50605. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50147 and NRRL B-50621 . In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50147 and NRRL B-50015. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50147 and NRRL B-50016. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50147 and NRRL B-50885. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50147 and NRRL B-50886. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50147 and NRRL B-50013. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50147 and NRRL B-50141. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50147 and NRRL B-50151 . In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50147 and NRRL B-50606. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50147 and PTA-7543. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50147 and PTA-7547. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50147 and NRRL B-50888. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50147 and PTA-7549. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50147 and NRRL B-50349.

In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50622 and NRRL B-50623. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50622 and NRRL B-50605. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50622 and NRRL B-50621. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50622 and NRRL B-50015. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50622 and NRRL B-50016. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50622 and NRRL B-50885. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50622 and NRRL B-50886. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50622 and NRRL B-50013. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50622 and NRRL B-50141 . In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50622 and NRRL B-50151. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50622 and NRRL B-50606. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50622 and PTA-7543. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50622 and PTA- 7547. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50622 and NRRL B-50888. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50622 and PTA-7549. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50622 and NRRL B-50349.

In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50623 and NRRL B-50605. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50623 and NRRL B-50621. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50623 and NRRL B-50015. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50623 and NRRL B-50016. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50623 and NRRL B-50885. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50623 and NRRL B-50886. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50623 and NRRL B-50013. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50623 and NRRL B-50141 . In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50623 and NRRL B-50151 . In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50623 and NRRL B-50606. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50623 and PTA-7543. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50623 and PTA-7547. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50623 and NRRL B-50888. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50623 and PTA-7549. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50623 and NRRL B-50349.

In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50605 and NRRL B-50621. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50605 and NRRL B-50015. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50605 and NRRL B-50016. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50605 and NRRL B-50885. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50605 and NRRL B-50886. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50605 and NRRL B-50013. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50605 and NRRL B-50141 . In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50605 and NRRL B-50151 . In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50605 and NRRL B-50606. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50605 and PTA-7543. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50605 and PTA-7547. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50605 and NRRL B-50888. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50605 and PTA-7549. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50605 and NRRL B-50349.

In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50621 and NRRL B-50015. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50621 and NRRL B-50016. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50621 and NRRL B-50885. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50621 and NRRL B-50886. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50621 and NRRL B-50013. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50621 and NRRL B-50141 . In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50621 and NRRL B-50151 . In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50621 and NRRL B-50606. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50621 and PTA-7543. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50621 and PTA-7547. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50621 and NRRL B-50888. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50621 and PTA-7549. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50621 and NRRL B-50349.

In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50015 and NRRL B-50016. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50015 and NRRL B-50885. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50015 and NRRL B-50886. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50015 and NRRL B-50013. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50015 and NRRL B-50141 . In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50015 and NRRL B-50151 . In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50015 and NRRL B-50606. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50015 and PTA-7543. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50015 and PTA-7547. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50015 and NRRL B-50888. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50015 and PTA-7549. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50015 and NRRL B- 50349.

In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50016 and NRRL B-50885. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50016 and NRRL B-50886. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50016 and NRRL B-50013. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50016 and NRRL B-50141. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50016 and NRRL B-50151. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50016 and NRRL B-50606. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50016 and PTA-7543. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50016 and PTA-7547. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50016 and NRRL B-50888. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50016 and PTA-7549. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50016 and NRRL B- 50349. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50885 and NRRL B-50886. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50885 and NRRL B-50013. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50885 and NRRL B-50141. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50885 and NRRL B-50151 . In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50885 and NRRL B-50606. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50885 and PTA-7543. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50885 and PTA-7547. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50885 and NRRL B-50888. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50885 and PTA-7549. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50885 and NRRL B-50349.

In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50886 and NRRL B-50013. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50886 and NRRL B-50141. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50886 and NRRL B-50151 . In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50886 and NRRL B-50606. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50886 and PTA-7543. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50886 and PTA- 7547. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50886 and NRRL B-50888. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50886 and PTA-7549. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50886 and NRRL B-50349. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50013 and NRRL B-50141. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50013 and NRRL B-50151 . In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50013 and NRRL B-50606. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50013 and PTA-7543. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50013 and PTA-7547. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50013 and NRRL B-50888. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50013 and PTA-7549. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50013 and NRRL B-50349.

In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50141 and NRRL B-50151 . In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50141 and NRRL B-50606. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50141 and PTA-7543. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50141 and PTA-7547. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50141 and NRRL B-50888. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50141 and PTA-7549. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50141 and NRRL B-50349.

In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50151 and NRRL B-50606. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50151 and PTA-7543. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50151 and PTA-7547. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50151 and NRRL B-50888. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50151 and PTA-7549. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50151 and NRRL B-50349.

In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50606 and PTA-7543. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50606 and PTA-7547. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50606 and NRRL B-50888. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50606 and PTA-7549. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50606 and NRRL B- 50349.

In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers PTA-7543 and PTA-7547. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers PTA-7543 and NRRL B-50888. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers PTA-7543 and PTA-7549. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers PTA-7543 and NRRL B-50349.

In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers PTA-7547 and NRRL B-50888. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers PTA-7547 and PTA-7549. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers PTA-7547 and NRRL B-50349.

In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50888 and PTA-7549. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers NRRL B-50888 and NRRL B-50349. In one embodiment, the method comprises adding to the animal feed the Bacillus strains having the deposit accession numbers PTA-7549 and NRRL B-50349.

In a preferred embodiment, the invention relates to a method for improving the digestibility of corn silage, comprising adding to the corn silage the Bacillus strain having the deposit accession number ATCC 700385 wherein the Bacillus strain improves the in vitro true digestibility of the corn silage. In a preferred embodiment, the invention relates to a method for improving the digestibility of corn silage, comprising adding to the corn silage the Bacillus strain having the deposit accession number NRRL B-50136 wherein the Bacillus strain improves the in vitro true digestibility of the corn silage.

In a preferred embodiment, the invention relates to a method for improving the digestibility of corn silage, comprising adding to the corn silage the Bacillus strain having the deposit accession number NRRL B-50147 wherein the Bacillus strain improves the in vitro true digestibility of the corn silage.

In a preferred embodiment, the invention relates to a method for improving the digestibility of corn silage, comprising adding to the corn silage the Bacillus strain having the deposit accession number NRRL B-50622 wherein the Bacillus strain improves the in vitro true digestibility of the corn silage.

In a preferred embodiment, the invention relates to a method for improving the digestibility of corn silage, comprising adding to the corn silage the Bacillus strain having the deposit accession number NRRL B-50623 wherein the Bacillus strain improves the in vitro true digestibility of the corn silage.

In a preferred embodiment, the invention relates to a method for improving the digestibility of corn silage, comprising adding to the corn silage the Bacillus strain having the deposit accession number NRRL B-50605 wherein the Bacillus strain improves the in vitro true digestibility of the corn silage.

In a preferred embodiment, the invention relates to a method for improving the digestibility of corn silage, comprising adding to the corn silage the Bacillus strain having the deposit accession number NRRL B-50621 wherein the Bacillus strain improves the in vitro true digestibility of the corn silage.

In a preferred embodiment, the invention relates to a method for improving the digestibility of corn silage, comprising adding to the corn silage the Bacillus strain having the deposit accession number PTA-7543 wherein the Bacillus strain improves the in vitro true digestibility of the corn silage.

In a preferred embodiment, the invention relates to a method for improving the digestibility of corn silage, comprising adding to the corn silage the Bacillus strain having the deposit accession number PTA-7547 wherein the Bacillus strain improves the in vitro true digestibility of the corn silage.

In a preferred embodiment, the invention relates to a method for improving the digestibility of corn silage, comprising adding to the corn silage the Bacillus strain having the deposit accession number NRRL B-50888 wherein the Bacillus strain improves the in vitro true digestibility of the corn silage. In a preferred embodiment, the invention relates to a method for improving the digestibility of corn silage, comprising adding to the corn silage the Bacillus strain having the deposit accession number PTA-7549 wherein the Bacillus strain improves the in vitro true digestibility of the corn silage.

In a preferred embodiment, the invention relates to a method for improving the digestibility of corn silage, comprising adding to the corn silage the Bacillus strain having the deposit accession number NRRL B-50349 wherein the Bacillus strain improves the in vitro true digestibility of the corn silage.

In a preferred embodiment, the invention relates to a method for improving the digestibility of corn silage, comprising adding to the corn silage the Bacillus strain having the deposit accession number NRRL B-50147 wherein the Bacillus strain improves the neutral detergent fiber digestibility of the corn silage.

In a preferred embodiment, the invention relates to a method for improving the digestibility of corn silage, comprising adding to the corn silage the Bacillus strain having the deposit accession number NRRL B-50622 wherein the Bacillus strain improves the neutral detergent fiber digestibility of the corn silage.

In a preferred embodiment, the invention relates to a method for improving the digestibility of corn silage, comprising adding to the corn silage the Bacillus strain having the deposit accession number NRRL B-50621 wherein the Bacillus strain improves the neutral detergent fiber digestibility of the corn silage.

In a preferred embodiment, the invention relates to a method for improving the digestibility of corn silage, comprising adding to the corn silage the Bacillus strain having the deposit accession number NRRL B-50141 wherein the Bacillus strain improves the neutral detergent fiber digestibility of the corn silage.

In a preferred embodiment, the invention relates to a method for improving the digestibility of corn silage, comprising adding to the corn silage the Bacillus strain having the deposit accession number NRRL B-50151 wherein the Bacillus strain improves the neutral detergent fiber digestibility of the corn silage.

In a preferred embodiment, the invention relates to a method for improving the digestibility of corn silage, comprising adding to the corn silage the Bacillus strain having the deposit accession number PTA-7543 wherein the Bacillus strain improves the neutral detergent fiber digestibility of the corn silage.

In a preferred embodiment, the invention relates to a method for improving the digestibility of corn silage, comprising adding to the corn silage the Bacillus strain having the deposit accession number PTA-7547 wherein the Bacillus strain improves the neutral detergent fiber digestibility of the corn silage. In a preferred embodiment, the invention relates to a method for improving the digestibility of corn silage, comprising adding to the corn silage the Bacillus strain having the deposit accession number NRRL B-50888 wherein the Bacillus strain improves the neutral detergent fiber digestibility of the corn silage.

In a preferred embodiment, the invention relates to a method for improving the digestibility of corn silage, comprising adding to the corn silage the Bacillus strain having the deposit accession number PTA-7549 wherein the Bacillus strain improves the neutral detergent fiber digestibility of the corn silage.

In a preferred embodiment, the invention relates to a method for improving the digestibility of corn silage, comprising adding to the corn silage the Bacillus strain having the deposit accession number NRRL B-50349 wherein the Bacillus strain improves the neutral detergent fiber digestibility of the corn silage.

In a preferred embodiment, the invention relates to a method for improving the digestibility of timothy hay, comprising adding to the timothy hay the Bacillus strain having the deposit accession number ATCC 700385 wherein the Bacillus strain improves the in vitro true digestibility of the timothy hay.

In a preferred embodiment, the invention relates to a method for improving the digestibility of timothy hay, comprising adding to the timothy hay the Bacillus strain having the deposit accession number NRRL B-50136 wherein the Bacillus strain improves the in vitro true digestibility of the timothy hay.

In a preferred embodiment, the invention relates to a method for improving the digestibility of timothy hay, comprising adding to the timothy hay the Bacillus strain having the deposit accession number NRRL B-50147 wherein the Bacillus strain improves the in vitro true digestibility of the timothy hay.

In a preferred embodiment, the invention relates to a method for improving the digestibility of timothy hay, comprising adding to the timothy hay the Bacillus strain having the deposit accession number NRRL B-50622 wherein the Bacillus strain improves the in vitro true digestibility of the timothy hay.

In a preferred embodiment, the invention relates to a method for improving the digestibility of timothy hay, comprising adding to the timothy hay the Bacillus strain having the deposit accession number NRRL B-50623 wherein the Bacillus strain improves the in vitro true digestibility of the timothy hay.

In a preferred embodiment, the invention relates to a method for improving the digestibility of timothy hay, comprising adding to the timothy hay the Bacillus strain having the deposit accession number NRRL B-50605 wherein the Bacillus strain improves the in vitro true digestibility of the timothy hay. In a preferred embodiment, the invention relates to a method for improving the digestibility of timothy hay, comprising adding to the timothy hay the Bacillus strain having the deposit accession number NRRL B-50621 wherein the Bacillus strain improves the in vitro true digestibility of the timothy hay.

In a preferred embodiment, the invention relates to a method for improving the digestibility of timothy hay, comprising adding to the timothy hay the Bacillus strain having the deposit accession number NRRL B-50015 wherein the Bacillus strain improves the in vitro true digestibility of the timothy hay.

In a preferred embodiment, the invention relates to a method for improving the digestibility of timothy hay, comprising adding to the timothy hay the Bacillus strain having the deposit accession number NRRL B-50016 wherein the Bacillus strain improves the in vitro true digestibility of the timothy hay.

In a preferred embodiment, the invention relates to a method for improving the digestibility of timothy hay, comprising adding to the timothy hay the Bacillus strain having the deposit accession number NRRL B-50885 wherein the Bacillus strain improves the in vitro true digestibility of the timothy hay.

In a preferred embodiment, the invention relates to a method for improving the digestibility of timothy hay, comprising adding to the timothy hay the Bacillus strain having the deposit accession number NRRL B-50886 wherein the Bacillus strain improves the in vitro true digestibility of the timothy hay.

In a preferred embodiment, the invention relates to a method for improving the digestibility of timothy hay, comprising adding to the timothy hay the Bacillus strain having the deposit accession number NRRL B-50013 wherein the Bacillus strain improves the in vitro true digestibility of the timothy hay.

In a preferred embodiment, the invention relates to a method for improving the digestibility of timothy hay, comprising adding to the timothy hay the Bacillus strain having the deposit accession number NRRL B-50141 wherein the Bacillus strain improves the in vitro true digestibility of the timothy hay.

In a preferred embodiment, the invention relates to a method for improving the digestibility of timothy hay, comprising adding to the timothy hay the Bacillus strain having the deposit accession number NRRL B-50151 wherein the Bacillus strain improves the in vitro true digestibility of the timothy hay.

In a preferred embodiment, the invention relates to a method for improving the digestibility of timothy hay, comprising adding to the timothy hay the Bacillus strain having the deposit accession number NRRL B-50606 wherein the Bacillus strain improves the in vitro true digestibility of the timothy hay. In a preferred embodiment, the invention relates to a method for improving the digestibility of timothy hay, comprising adding to the timothy hay the Bacillus strain having the deposit accession number PTA-7543 wherein the Bacillus strain improves the in vitro true digestibility of the timothy hay.

In a preferred embodiment, the invention relates to a method for improving the digestibility of timothy hay, comprising adding to the timothy hay the Bacillus strain having the deposit accession number PTA-7547 wherein the Bacillus strain improves the in vitro true digestibility of the timothy hay.

In a preferred embodiment, the invention relates to a method for improving the digestibility of timothy hay, comprising adding to the timothy hay the Bacillus strain having the deposit accession number NRRL B-50888 wherein the Bacillus strain improves the in vitro true digestibility of the timothy hay.

In a preferred embodiment, the invention relates to a method for improving the digestibility of timothy hay, comprising adding to the timothy hay the Bacillus strain having the deposit accession number PTA-7549 wherein the Bacillus strain improves the in vitro true digestibility of the timothy hay.

In a preferred embodiment, the invention relates to a method for improving the digestibility of timothy hay, comprising adding to the timothy hay the Bacillus strain having the deposit accession number NRRL B-50349 wherein the Bacillus strain improves the in vitro true digestibility of the timothy hay.

In a preferred embodiment, the invention relates to a method for improving the digestibility of timothy hay, comprising adding to the timothy hay the Bacillus strain having the deposit accession number ATCC 700385 wherein the Bacillus strain improves the neutral detergent fiber digestibility of the timothy hay.

In a preferred embodiment, the invention relates to a method for improving the digestibility of timothy hay, comprising adding to the timothy hay the Bacillus strain having the deposit accession number NRRL B-50147 wherein the Bacillus strain improves the neutral detergent fiber digestibility of the timothy hay.

In a preferred embodiment, the invention relates to a method for improving the digestibility of timothy hay, comprising adding to the timothy hay the Bacillus strain having the deposit accession number NRRL B-50622 wherein the Bacillus strain improves the neutral detergent fiber digestibility of the timothy hay.

In a preferred embodiment, the invention relates to a method for improving the digestibility of timothy hay, comprising adding to the timothy hay the Bacillus strain having the deposit accession number NRRL B-50623 wherein the Bacillus strain improves the neutral detergent fiber digestibility of the timothy hay. In a preferred embodiment, the invention relates to a method for improving the digestibility of timothy hay, comprising adding to the timothy hay the Bacillus strain having the deposit accession number NRRL B-50605 wherein the Bacillus strain improves the neutral detergent fiber digestibility of the timothy hay.

In a preferred embodiment, the invention relates to a method for improving the digestibility of timothy hay, comprising adding to the timothy hay the Bacillus strain having the deposit accession number NRRL B-50621 wherein the Bacillus strain improves the neutral detergent fiber digestibility of the timothy hay.

In a preferred embodiment, the invention relates to a method for improving the digestibility of timothy hay, comprising adding to the timothy hay the Bacillus strain having the deposit accession number NRRL B-50015 wherein the Bacillus strain improves the neutral detergent fiber digestibility of the timothy hay.

In a preferred embodiment, the invention relates to a method for improving the digestibility of timothy hay, comprising adding to the timothy hay the Bacillus strain having the deposit accession number NRRL B-50885 wherein the Bacillus strain improves the neutral detergent fiber digestibility of the timothy hay.

In a preferred embodiment, the invention relates to a method for improving the digestibility of timothy hay, comprising adding to the timothy hay the Bacillus strain having the deposit accession number NRRL B-50886 wherein the Bacillus strain improves the neutral detergent fiber digestibility of the timothy hay.

In a preferred embodiment, the invention relates to a method for improving the digestibility of timothy hay, comprising adding to the timothy hay the Bacillus strain having the deposit accession number NRRL B-50013 wherein the Bacillus strain improves the neutral detergent fiber digestibility of the timothy hay.

In a preferred embodiment, the invention relates to a method for improving the digestibility of timothy hay, comprising adding to the timothy hay the Bacillus strain having the deposit accession number NRRL B-50141 wherein the Bacillus strain improves the neutral detergent fiber digestibility of the timothy hay.

In a preferred embodiment, the invention relates to a method for improving the digestibility of timothy hay, comprising adding to the timothy hay the Bacillus strain having the deposit accession number NRRL B-50151 wherein the Bacillus strain improves the neutral detergent fiber digestibility of the timothy hay.

In a preferred embodiment, the invention relates to a method for improving the digestibility of timothy hay, comprising adding to the timothy hay the Bacillus strain having the deposit accession number NRRL B-50606 wherein the Bacillus strain improves the neutral detergent fiber digestibility of the timothy hay. In a preferred embodiment, the invention relates to a method for improving the digestibility of timothy hay, comprising adding to the timothy hay the Bacillus strain having the deposit accession number PTA-7543 wherein the Bacillus strain improves the neutral detergent fiber digestibility of the timothy hay.

In a preferred embodiment, the invention relates to a method for improving the digestibility of timothy hay, comprising adding to the timothy hay the Bacillus strain having the deposit accession number PTA-7547 wherein the Bacillus strain improves the neutral detergent fiber digestibility of the timothy hay.

In a preferred embodiment, the invention relates to a method for improving the digestibility of timothy hay, comprising adding to the timothy hay the Bacillus strain having the deposit accession number NRRL B-50888 wherein the Bacillus strain improves the neutral detergent fiber digestibility of the timothy hay.

In a preferred embodiment, the invention relates to a method for improving the digestibility of timothy hay, comprising adding to the timothy hay the Bacillus strain having the deposit accession number PTA-7549 wherein the Bacillus strain improves the neutral detergent fiber digestibility of the timothy hay.

In a preferred embodiment, the invention relates to a method for improving the digestibility of timothy hay, comprising adding to the timothy hay the Bacillus strain having the deposit accession number NRRL B-50349 wherein the Bacillus strain improves the neutral detergent fiber digestibility of the timothy hay.

In a more preferred embodiment of the method, the animal feed is fed to a ruminant. In an even more embodiment of the method, the ruminant is a cattle, cow, beef cattle, young calf, goat, sheep, lamb, deer, yank, camel or llama. In another embodiment of the method, the animal feed further comprises one or more components selected from the list consisting of forage; concentrate; one or more enzymes; one or more additional microbes; one or more vitamins; one or more minerals; one or more amino acids; and one or more other feed ingredients.

In another embodiment of the method, the animal feed further comprises forage. In another embodiment of the method, the animal feed further comprises concentrate. In another embodiment of the method, the animal feed further comprises one or more additional microbes. In another embodiment of the method, the animal feed further comprises one or more enzymes. In another embodiment of the method, the animal feed further comprises one or more vitamins. In another embodiment of the method, the animal feed further comprises one or more minerals. In another embodiment of the method, the animal feed further comprises one or more amino acids. In another embodiment of the method, the animal feed further comprises one or more other feed ingredients. In an embodiment to any of the aforementioned embodiments, the method also improves the digestibility of the animal feed, such as when the animal feed is sub-optimal. In another embodiment to any of the aforementioned embodiments, the method also increases the milk yield of the ruminant. In an embodiment to any of the aforementioned embodiments, the method also increases the meat yield of the ruminant.

In still yet another embodiment of the method, the one or more bacterial strains are present in the form of a stable spore. In still a further embodiment of the method, the stable spore will germinate in the rumen of the ruminant.

In a preferred embodiment, the method comprises administering to a ruminant one or more bacterial strains described herein, wherein the bacterial count of each of the bacterial strains is between 1 x10 4 and 1x10 14 CFU/kg of forage, preferably between 1 x10 6 and 1 x10 12 CFU/kg of forage, and more preferably between 1 x10 7 and 1 x10 11 CFU/kg of forage. In a more preferred embodiment the bacterial count of each of the bacterial strains described herein is between 1x10 8 and 1x10 10 CFU/kg of forage.

In a preferred embodiment, the method comprises administering to a ruminant one or more bacterial strains described herein, wherein the bacterial count of each of the bacterial strains is between 1x10 5 and 1 x10 15 CFU/animal/day, preferably between 1x10 7 and 1 x10 13 CFU/animal/day, and more preferably between 1x10 8 and 1 x10 12 CFU/animal/day. In a more preferred embodiment the bacterial count of each of the bacterial strains described herein is between 1x10 9 and 1 x10 11 CFU/animal/day.

In one aspect the invention relates to a method for improving the digestibility of an animal feed such as a ruminant feed comprising:

(a) feeding an animal such as a ruminant a feed; and

(b) administering to an animal such as a ruminant a Bacillus strain selected from the group consisting of Bacillus pumilus, Bacillus amyloliquefaciens, Bacillus subtilis and

Bacillus licheniformis or any combination thereof wherein the Bacillus strain improves the in vitro true digestibility and/or neutral detergent fiber digestibility and/or the Acid Detergent Fiber Digestibility (ADFD) of the animal feed and wherein the Bacillus strain has enzyme activity against Casein and/or Xylan under aerobic conditions.

In another aspect, the invention covers the method for improving the digestibility of an animal feed comprising:

(a) feeding an animal such as a ruminant a feed; and

(b) administering to an animal such as a ruminant one or more Bacillus strains selected from the group consisting of:

the strain having the deposit accession number ATCC 700385; the strain having the deposit accession number NRRL B-50136; the strain having the deposit accession number NRRL B-50147; the strain having the deposit accession number NRRL B-50622; the strain having the deposit accession number NRRL B-50623; the strain having the deposit accession number NRRL B-50605; the strain having the deposit accession number NRRL B-50621 ; the strain having the deposit accession number NRRL B-50015; the strain having the deposit accession number NRRL B-50016; the strain having the deposit accession number NRRL B-50885; the strain having the deposit accession number NRRL B-50886; the strain having the deposit accession number NRRL B-50013; the strain having the deposit accession number NRRL B-50141 ; the strain having the deposit accession number NRRL B-50151 ; the strain having the deposit accession number NRRL B-50606; the strain having the deposit accession number PTA-7543;

the strain having the deposit accession number PTA-7547;

the strain having the deposit accession number NRRL B-50888; the strain having the deposit accession number PTA-7549; and the strain having the deposit accession number NRRL B-50349, or any combination thereof,

wherein the Bacillus strain improves the in vitro true digestibility and/or neutral detergent fiber digestibility and/or the Acid Detergent Fiber Digestibility (ADFD) of the animal feed, and wherein step (a) occurs before, after, or simultaneously with step (b).

In a further embodiment, the method comprises administering to the animal feed one or more bacterial strains such as at least two of the above strains, at least three of the above strains, at least four of the above strains, at least five of the above strains, at least six of the above strains, at least seven of the above strains, at least eight of the above strains, at least nine of the above strains, at least ten of the above strains, at least eleven of the above strains, at least twelve of the above strains, at least thirteen of the above strains, at least fourteen of the above strains, at least fifteen of the above strains, at least sixteen of the above strains, at least seventeen of the above strains, at least eighteen of the above strains, at least nineteen of the above strains, up to and including all of the above strains.

In a preferred embodiment of the method, the animal feed is fed to a ruminant. In an embodiment of the method, the ruminant is a cattle, cow, beef cattle, young calf, goat, sheep, lamb, deer, yank, camel or llama. In another embodiment of the method, the animal feed further comprises one or more components selected from the list consisting of forage; concentrate; one or more enzymes; one or more additional microbes; one or more vitamins; one or more minerals; one or more amino acids; and one or more other feed ingredients. In another embodiment of the method, the animal feed further comprises forage. In another embodiment of the method, the animal feed further comprises concentrate. In another embodiment of the method, the animal feed further comprises one or more additional microbes. In another embodiment of the method, the animal feed further comprises one or more enzymes. In another embodiment of the method, the animal feed further comprises one or more vitamins. In another embodiment of the method, the animal feed further comprises one or more minerals. In another embodiment of the method, the animal feed further comprises one or more amino acids. In another embodiment of the method, the animal feed further comprises one or more other feed ingredients.

In an embodiment to any of the aforementioned embodiments, the method also improves the digestibility of the animal feed, such as when the animal feed is sub-optimal. In another embodiment to any of the aforementioned embodiments, the method also increases the milk yield of the ruminant. In an embodiment to any of the aforementioned embodiments, the method also increases the meat yield of the ruminant.

In still yet another embodiment of the method, the one or more bacterial strains are present in the form of a stable spore. In still a further embodiment of the method, the stable spore will germinate in the rumen of the ruminant.

In a preferred embodiment, the method comprises administering to a ruminant one or more bacterial strains described herein, wherein the bacterial count of each of the bacterial strains is between 1 x10 4 and 1x10 14 CFU/kg of forage, preferably between 1 x10 6 and 1 x10 12 CFU/kg of forage, and more preferably between 1 x10 7 and 1 x10 11 CFU/kg of forage. In a more preferred embodiment the bacterial count of each of the bacterial strains described herein is between 1x10 8 and 1x10 10 CFU/kg of forage.

In a preferred embodiment, the method comprises administering to a ruminant one or more bacterial strains described herein, wherein the bacterial count of each of the bacterial strains is between 1x10 5 and 1 x10 15 CFU/animal/day, preferably between 1x10 7 and 1 x10 13 CFU/animal/day, and more preferably between 1x10 8 and 1 x10 12 CFU/animal/day. In a more preferred embodiment the bacterial count of each of the bacterial strains described herein is between 1x10 9 and 1 x10 11 CFU/animal/day.

Strains

In a preferred embodiment the invention relates to a Bacillus strain selected from the group consisting of:

the strain having the deposit accession number PTA-7541 or a strain having all of the identifying characteristics of Bacillus PTA-7541 or a mutant thereof;

the strain having the deposit accession number NRRL B-676141 or a strain having all of the identifying characteristics of Bacillus NRRL B-676141 or a mutant thereof; and the strain having the deposit accession number NRRL B-676140 or a strain having all of the identifying characteristics of Bacillus NRRL B-676140 or a mutant thereof.

Compositions

In another aspect, the invention relates to a composition such as an animal feed composition such as a ruminant feed comprising a carrier, such as forage and one or more of the bacteria cultures having characteristics substantially identical to that of a strain selected from the group consisting of:

the strain having the deposit accession number NRRL B-50885 or a strain having all of the identifying characteristics of Bacillus NRRL B-50885 or a mutant thereof;

the strain having the deposit accession number NRRL B-50886 or a strain having all of the identifying characteristics of Bacillus NRRL B-50886 or a mutant thereof;

the strain having the deposit accession number NRRL B-50888 or a strain having all of the identifying characteristics of Bacillus NRRL B-50888 or a mutant thereof,

the strain having the deposit accession number PTA-7541 or a strain having all of the identifying characteristics of Bacillus PTA-7541 or a mutant thereof;

the strain having the deposit accession number NRRL B-59648 or a strain having all of the identifying characteristics of Bacillus NRRL B-59648 or a mutant thereof;

the strain having the deposit accession number NRRL B-59650 or a strain having all of the identifying characteristics of Bacillus NRRL B-59650 or a mutant thereof;

the strain having the deposit accession number NRRL B-59651 or a strain having all of the identifying characteristics of Bacillus NRRL B-59651 or a mutant thereof;

the strain having the deposit accession number NRRL B-676141 or a strain having all of the identifying characteristics of Bacillus NRRL B-676141 or a mutant thereof; and

the strain having the deposit accession number NRRL B-676140 or a strain having all of the identifying characteristics of Bacillus NRRL B-676140 or a mutant thereof,

or any combination thereof.

In an embodiment, the animal feed composition comprises forage and the strain having the deposit accession number NRRL B-50885. In an embodiment, the animal feed composition comprises forage and the strain having the deposit accession number NRRL B- 50886. In an embodiment, the animal feed composition comprises forage and the strain having the deposit accession number NRRL B-50888. In an embodiment, the animal feed composition comprises forage and the strain having the deposit accession number PTA- 7541. In an embodiment, the animal feed composition comprises forage and the strain having the deposit accession number NRRL B-59648. In an embodiment, the animal feed composition comprises forage and the strain having the deposit accession number B-59650. In an embodiment, the animal feed composition comprises forage and the strain having the deposit accession number NRRL B-59651 . In an embodiment, the animal feed composition comprises forage and the strain having the deposit accession number NRRL B-676141. In an embodiment, the animal feed composition comprises forage and the strain having the deposit accession number NRRL B-676140.

In an embodiment, the animal feed composition comprises at least two or more bacteria cultures, at least three or more bacteria cultures, at least four or more bacteria cultures, or all of the bacterial cultures of the invention.

In an embodiment, the animal feed composition comprises the bacteria cultures having characteristics substantially identical to that of the strain having the deposit accession number NRRL B-50885 and the strain having the deposit accession number NRRL B-50886.

In an embodiment, the animal feed composition comprises the bacteria cultures having characteristics substantially identical to that of the strain having the deposit accession number NRRL B-50885 and the strain having the deposit accession number NRRL B-50888.

In an embodiment, the animal feed composition comprises the bacteria cultures having characteristics substantially identical to that of the strain having the deposit accession number NRRL B-50886 and the strain having the deposit accession number NRRL B-50888.

In another embodiment, the animal feed composition is for feeding to a ruminant, such as cattle, cow, beef cattle, young calf, goat, sheep, lamb, deer, yank, camel or llama. In another embodiment, the animal feed further comprises one or more components selected from the list consisting of forage; concentrate; one or more enzymes; one or more additional microbes; one or more vitamins; one or more minerals; one or more amino acids; and one or more other feed ingredients as described below.

In a preferred embodiment, the bacterial count of each of the bacterial strains in the animal feed composition is between 1 x10 4 and 1x10 14 CFU/kg of dry matter, preferably between 1 x10 6 and 1 x10 12 CFU/kg of dry matter, and more preferably between 1 x10 7 and

1x10 11 CFU/kg of dry matter. In a more preferred embodiment the bacterial count of each of the bacterial strains in the animal feed composition is between 1 x10 8 and 1 x10 10 CFU/kg of dry matter.

In a preferred embodiment, the bacterial count of each of the bacterial strains in the animal feed composition is between 1x10 5 and 1 x10 15 CFU/animal/day, preferably between 1x10 7 and 1 x10 13 CFU/animal/day, and more preferably between 1 x10 8 and 1x10 12 CFU/animal/day. In a more preferred embodiment the bacterial count of each of the bacterial strains in the animal feed composition is between 1 x10 9 and 1x10 11 CFU/animal/day.

In still yet another embodiment, the one or more bacterial strains are present in the form of a stable spore. In still a further embodiment, the stable spore will germinate in the rumen of the ruminant. In one embodiment, the one or more bacterial strains are stable when subjected to pressures applied/achieved during an extrusion process for pelleting. In a particular embodiment, the one or more bacterial strains are stable at pressures ranging from 1 bar to 40 bar, particularly 10 bar to 40 bar, more particularly 15 bar to 40 bar, even more particularly 20 bar to 40 bar, still even more particularly 35 bar to 37 bar, even still more particularly 36 bar.

In a particular embodiment, the one or more bacterial strains are stable at high temperatures. In particular, the bacterial strains are stable when they are subjected to temperatures achieved during an extrusion process for pelleting. In an even more particular embodiment, the one or more bacterial strains are stable at temperatures ranging from 80°C to 120°C, particularly temperatures ranging from, 90°C to 120°C, even more particularly temperatures ranging from 95°C to 120°C.

In an embodiment, the animal feed composition further comprises one or more additional microbes. In a particular embodiment, the animal feed composition further comprises a bacterium from one or more of the following genera: Lactobacillus, Lactococcus, Streptococcus, Bacillus, Pediococcus, Enterococcus, Leuconostoc, Carnobacterium, Propionibacterium, Bifidobacterium, Clostridium and Megasphaera or any combination thereof.

In a particular embodiment, animal feed composition further comprises a bacterium from one or more of the following strains of Bacillus amyloliquifaciens, Bacillus subtilis, Bacillus pumilus, Bacillus polymyxa, Bacillus licheniformis, Bacillus megaterium, Bacillus coagulans, Bacillus circulans, or any combination thereof.

In a particular embodiment, the animal feed composition further comprises one or more types of yeast. The one or more types of yeast can be selected from the group consisting of Saccharomycetaceae, Saccharomyces (such as S. cerevisiae and/or S. boulardii), Kluyveromyces (such as K. marxianus and K. lactis), Candida (such as C. utilis, also called Torula yeast), Pichia (such as P. pastoris), Torulaspora (such as T. delbrueckii), Phaffia yeasts and Basidiomycota.

In a more particular embodiment, the animal feed composition further comprises one or more bacterium from one or more Bacillus strains selected from the group consisting of: the strain having the deposit accession number ATCC 700385;

the strain having the deposit accession number NRRL B-50623;

the strain having the deposit accession number NRRL B-50621 ;

the strain having the deposit accession number NRRL B-50015;

the strain having the deposit accession number NRRL B-50013;

the strain having the deposit accession number PTA-7543;

the strain having the deposit accession number PTA-7549; the strain having the deposit accession number PTA-7541 ;

the strain having the deposit accession number NRRL B-59648;

the strain having the deposit accession number NRRL B-59650;

the strain having the deposit accession number NRRL B-59651 ;

the strain having the deposit accession number NRRL B-676141 ; and

the strain having the deposit accession number NRRL B-676140

or any combination thereof, such as, at least three of the above strains, at least four of the above strains, at least five of the above strains, at least six of the above strains up to and including all of the above strains.

In a preferred embodiment, the bacterial count of each of the bacterial strains in the animal feed composition is between 1 x10 4 and 1x10 14 CFU/kg of dry matter, preferably between 1 x10 6 and 1 x10 12 CFU/kg of dry matter, and more preferably between 1 x10 7 and 1x10 11 CFU/kg of dry matter. In a more preferred embodiment the bacterial count of each of the bacterial strains in the animal feed composition is between 1 x10 8 and 1 x10 10 CFU/kg of dry matter.

In a preferred embodiment, the bacterial count of each of the bacterial strains in the animal feed composition is between 1x10 5 and 1 x10 15 CFU/animal/day, preferably between 1x10 7 and 1 x10 13 CFU/animal/day, and more preferably between 1 x10 8 and 1x10 12 CFU/animal/day. In a more preferred embodiment the bacterial count of each of the bacterial strains in the animal feed composition is between 1 x10 9 and 1x10 11 CFU/animal/day.

In still yet another embodiment, the one or more bacterial strains are present in the form of a stable spore. In still a further embodiment, the stable spore will germinate in the rumen of the ruminant. Animal Feed

In one aspect, the animal feed, comprises forage and may further comprise concentrates as well as vitamins, minerals, enzymes, amino acids and/or other feed ingredients (such as in a premix).

Forage as defined herein also includes roughage. Forage is fresh plant material such as hay and silage from forage plants, grass and other forage plants, grass and other forage plants, seaweed, sprouted grains and legumes, or any combination thereof. Examples of forage plants are Alfalfa (lucerne), birdsfoot trefoil, brassica (e.g. kale, rapeseed (canola), rutabaga (swede), turnip), clover (e.g. alsike clover, red clover, subterranean clover, white clover), grass (e.g. Bermuda grass, brome, false oat grass, fescue, heath grass, meadow grasses, orchard grass, ryegrass, Timothy-grass), corn (maize), millet, barley, oats, rye, sorghum, soybeans and wheat and vegetables such as beets. Crops suitable for ensilage are the ordinary grasses, clovers, alfalfa, vetches, oats, rye and maize. Forage further includes crop residues from grain production (such as corn stover; straw from wheat, barley, oat, rye and other grains); residues from vegetables like beet tops; residues from oilseed production like stems and leaves form soy beans, rapeseed and other legumes; and fractions from the refining of grains for animal or human consumption or from fuel production or other industries.

Roughage is generally dry plant material with high levels of fiber, such as fiber, bran, husks from seeds and grains and crop residues (such as stover, copra, straw, chaff, sugar beet waste).

Examples of concentrates are feed with high protein and energy concentrations, such as fish meal, molasses, oligosaccharides, sorghum, seeds and grains (either whole or prepared by crushing, milling, etc from e.g. corn, oats, rye, barley, wheat), oilseed press cake (e.g. from cottonseed, safflower, sunflower, soybean, rapeseed/canola, peanut or groundnut), palm kernel cake, yeast derived material and distillers grains (such as wet distillers grains (WDS) and dried distillers grains with solubles (DDGS)).

In one embodiment, the forage and one or more microbes are mixed with a concentrate. In another embodiment, the forage and one or more microbes are mixed with a premix. In a further embodiment, the forage and one or more microbes are mixed with vitamins and/or minerals. In a further embodiment, the forage and one or more microbes are mixed with one or more enzymes. In a further embodiment, the forage and one or more microbes are mixed with other feed ingredients, such as colouring agents, stabilisers, growth improving additives and aroma compounds/flavorings, polyunsaturated fatty acids (PUFAs); reactive oxygen generating species, anti-microbial peptides, anti-fungal polypeptides and amino acids.

In particular embodiments, the animal feed may comprise 0-80% maize; and/or 0- 80% sorghum; and/or 0-70% wheat; and/or 0-70% barley; and/or 0-30% oats; and/or 0-40% soybean meal; and/or 0-10% fish meal; and/or 0-20% whey.

The animal feed may comprise vegetable proteins. In particular embodiments, the protein content of the vegetable proteins is at least 10, 20, 30, 40, 50, 60, 70, 80, or 90% (w/w). Vegetable proteins may be derived from vegetable protein sources, such as legumes and cereals, for example, materials from plants of the families Fabaceae (Leguminosae), Cruciferaceae, Chenopodiaceae, and Poaceae, such as soy bean meal, lupin meal, rapeseed meal, and combinations thereof.

In a particular embodiment, the vegetable protein source is material from one or more plants of the family Fabaceae, e.g., soybean, lupine, pea, or bean. In another particular embodiment, the vegetable protein source is material from one or more plants of the family Chenopodiaceae, e.g. beet, sugar beet, spinach or quinoa. Other examples of vegetable protein sources are rapeseed, and cabbage. In another particular embodiment, soybean is a preferred vegetable protein source. Other examples of vegetable protein sources are cereals such as barley, wheat, rye, oat, maize (corn), rice, and sorghum.

In a preferred embodiment the invention relates to a ruminant feed with a 48 hour in vitro NDF Digestibility of 30-70% of NDF such as a 48 hour in vitro NDF Digestibility of NDF selected from the group consisting of from 30-40%, for example from 40-50%, such as from 50-60%, for example from 60-70%, or any combination of these intervals (as defined in "Using NDF Digestibility in Ration Formulation, Hoffman and Combs, Focus on Forage Vol. 6: No. 3). Premix

In an embodiment, the animal feed may include a premix, comprising e.g. vitamins, minerals, enzymes, amino acids, preservatives, antibiotics, other feed ingredients or any combination thereof which are mixed into the animal feed. Vitamins and Minerals

In another embodiment, the animal feed may include one or more vitamins, such as one or more fat-soluble vitamins and/or one or more water-soluble vitamins. In another embodiment, the animal feed may optionally include one or more minerals, such as one or more trace minerals and/or one or more macro minerals.

Usually fat- and water-soluble vitamins, as well as trace minerals form part of a so- called premix intended for addition to the feed, whereas macro minerals are usually separately added to the feed.

Non-limiting examples of fat-soluble vitamins include vitamin A, vitamin D3, vitamin E, and vitamin K, e.g., vitamin K3.

Non-limiting examples of water-soluble vitamins include vitamin B12, biotin and choline, vitamin B1 , vitamin B2, vitamin B6, niacin, folic acid and panthothenate, e.g., Ca-D- panthothenate.

Non-limiting examples of trace minerals include boron, cobalt, chloride, chromium, copper, fluoride, iodine, iron, manganese, molybdenum, selenium and zinc.

Non-limiting examples of macro minerals include calcium, magnesium, potassium and sodium.

Enzymes

In another embodiment, the animal feed compositions described herein optionally include one or more enzymes. Enzymes can be classified on the basis of the handbook Enzyme Nomenclature from NC-IUBMB, 1992), see also the ENZYME site at the internet: www.expasy.ch/enzyme/. ENZYME is a repository of information relative to the nomenclature of enzymes. It is primarily based on the recommendations of the Nomenclature Committee of the International Union of Biochemistry and Molecular Biology (IUB-MB), Academic Press, Inc., 1992, and it describes each type of characterized enzyme for which an EC (Enzyme Commission) number has been provided (Bairoch, 2000, The ENZYME database, Nucleic Acids Res 28: 304-305). This IUB-MB Enzyme nomenclature is based on their substrate specificity and occasionally on their molecular mechanism; such a classification does not reflect the structural features of these enzymes.

Another classification of certain glycoside hydrolase enzymes, such as endoglucanase, xylanase, galactanase, mannanase, dextranase and alpha-galactosidase, in families based on amino acid sequence similarities has been proposed a few years ago. They currently fall into 90 different families: See the CAZy(ModO) internet site (Coutinho, P.M. & Henrissat, B. (1999) Carbohydrate-Active Enzymes server at URL: afmb.cnrs- mrs.fr/~cazy/CAZY/index.html (corresponding papers: Coutinho, P.M. & Henrissat, B. (1999) Carbohydrate-active enzymes: an integrated database approach. In "Recent Advances in Carbohydrate Bioengineering", H.J. Gilbert, G. Davies, B. Henrissat and B. Svensson eds., The Royal Society of Chemistry, Cambridge, pp. 3-12; Coutinho, P.M. & Henrissat, B. (1999) The modular structure of cellulases and other carbohydrate-active enzymes: an integrated database approach. In "Genetics, Biochemistry and Ecology of Cellulose Degradation"., K. Ohmiya, K. Hayashi, K. Sakka, Y. Kobayashi, S. Karita and T. Kimura eds., Uni Publishers Co., Tokyo, pp. 15-23).

Thus the composition of the invention may also comprise at least one other enzyme selected from the group comprising of phytase (EC 3.1 .3.8 or 3.1.3.26); xylanase (EC 3.2.1.8); galactanase (EC 3.2.1 .89); alpha-galactosidase (EC 3.2.1.22); protease (EC 3.4); phospholipase A1 (EC 3.1 .1.32); phospholipase A2 (EC 3.1.1.4); lysophospholipase (EC 3.1 .1.5); phospholipase C (3.1.4.3); phospholipase D (EC 3.1.4.4); amylase such as, for example, alpha-amylase (EC 3.2.1.1 ); lysozyme (EC 3.2.1 .17); and beta-glucanase (EC 3.2.1.4 or EC 3.2.1.6), or any mixture thereof.

In a particular embodiment, the composition of the invention comprises a phytase (EC 3.1 .3.8 or 3.1.3.26). Examples of commercially available phytases include Bio-Feed™ Phytase (Novozymes), Ronozyme® P and HiPhos™ (DSM Nutritional Products), Natuphos™ (BASF), Finase® and Quantum® Blue (AB Enzymes), the Phyzyme® XP (Verenium/DuPont) and Axtra® PHY (DuPont). Other preferred phytases include those described in e.g. WO 98/28408, WO 00/43503, and WO 03/066847.

In a particular embodiment, the composition of the invention comprises a xylanase (EC 3.2.1.8). Examples of commercially available xylanases include Ronozyme® WX and G2 (DSM Nutritional Products), Econase® XT and Barley (AB Vista), Xylathin® (Verenium) and Axtra® XB (Xylanase/beta-glucanase, DuPont) In a particular embodiment, the composition of the invention comprises a protease (EC 3.4). Examples of commercially available proteases include Ronozyme® ProAct (DSM Nutritional Products). Amino Acids

The composition of the invention may further comprise one or more amino acids. Examples of amino acids which are used in animal feed are lysine, alanine, beta-alanine, threonine, methionine and tryptophan. Other feed ingredients

The composition of the invention may further comprise colouring agents, stabilisers, growth improving additives and aroma compounds/flavorings, polyunsaturated fatty acids (PUFAs); reactive oxygen generating species, anti-microbial peptides and anti-fungal polypeptides.

Examples of colouring agents are carotenoids such as beta-carotene, astaxanthin, and lutein.

Examples of aroma compounds/flavorings are creosol, anethol, deca-, undeca-and/or dodeca-lactones, ionones, irone, gingerol, piperidine, propylidene phatalide, butylidene phatalide, capsaicin and tannin.

Examples of antimicrobial peptides (AMP's) are CAP18, Leucocin A, Tritrpticin,

Protegrin-1 , Thanatin, Defensin, Lactoferrin, Lactoferricin, and Ovispirin such as Novispirin (Robert Lehrer, 2000), Plectasins, and Statins, including the compounds and polypeptides disclosed in WO 03/044049 and WO 03/048148, as well as variants or fragments of the above that retain antimicrobial activity.

Examples of antifungal polypeptides (AFP's) are the Aspergillus giganteus, and

Aspergillus niger peptides, as well as variants and fragments thereof which retain antifungal activity, as disclosed in WO 94/01459 and WO 02/090384.

Examples of polyunsaturated fatty acids are C18, C20 and C22 polyunsaturated fatty acids, such as arachidonic acid, docosohexaenoic acid, eicosapentaenoic acid and gamma- linoleic acid.

Examples of reactive oxygen generating species are chemicals such as perborate, persulphate, or percarbonate; and enzymes such as an oxidase, an oxygenase or a syntethase.

The composition of the invention may further comprise at least one amino acid. Examples of amino acids which are used in animal feed are lysine, alanine, beta-alanine, threonine, methionine and tryptophan. Manufacturing

Animal diets can e.g. be manufactured as mash feed (non-pelleted) or pelleted feed. Typically, the milled feed-stuffs are mixed and sufficient amounts of essential vitamins and minerals are added according to the specifications for the species in question. The bacteria cultures and optionally enzymes can be added as solid or liquid formulations. For example, for mash feed a solid or liquid culture formulation may be added before or during the ingredient mixing step. For pelleted feed the (liquid or solid) culture preparation may also be added before or during the feed ingredient step. Typically a liquid culture preparation comprises the culture of the invention optionally with a polyol, such as glycerol, ethylene glycol or propylene glycol, and is added after the pelleting step, such as by spraying the liquid formulation onto the pellets. The enzyme may also be incorporated in a feed additive or premix.

The enzyme may be added to the feed mix as a granule, which is optionally pelleted or extruded. The granule typically comprises a core particle and one or more coatings, which typically are salt and/or wax coatings. The core particle can either be a homogeneous blend of an active compound optionally together with salts (e.g. organic or inorganic zinc or calcium salt) or an inert particle with an active compound applied onto it. The active compound is the culture of the invention optionally combined with one or more enzymes. The inert particle may be water soluble or water insoluble, e.g. starch, a sugar (such as sucrose or lactose), or a salt (such as NaCI, Na 2 S0 4 ). The salt coating is typically at least 1 μηη thick and can either be one particular salt or a mixture of salts, such as Na 2 S0 4 , K 2 S0 4 , MgS0 4 and/or sodium citrate. Other examples are those described in e.g. WO 2008/017659, WO 2006/034710, WO 97/05245, WO 98/54980, WO 98/55599, WO 00/70034 or polymer coating such as described in WO 01/00042.

Alternatively, the protease can be prepared by freezing a mixture of liquid culture solution with a bulking agent such as ground soybean meal, and then lyophilizing the mixture.

Preferred Embodiments

Preferred embodiments of the invention are described in the set of items herein below.

1. A method for improving the digestibility of an animal feed such as a ruminant feed, comprising adding to the animal feed a Bacillus strain selected from the group consisting of Bacillus pumilus, Bacillus amyloliquefaciens, Bacillus subtilis and Bacillus licheniformis wherein the Bacillus strain improves the in vitro true digestibility and/or neutral detergent fiber digestibility of the animal feed and wherein the Bacillus strain has enzyme activity against Casein or Xylan under aerobic conditions e.g. as determined as described in Example 1 to 4.

2. The method according to item 1 comprising adding to the animal feed a Bacillus strain selected from the group consisting of:

the strain having the deposit accession number ATCC 700385 or a strain having all of the identifying characteristics of Bacillus ATCC 700385 or a mutant thereof;

the strain having the deposit accession number NRRL B-50136 or a strain having all of the identifying characteristics of Bacillus NRRL B-50136 or a mutant thereof;

the strain having the deposit accession number NRRL B-50147 or a strain having all of the identifying characteristics of Bacillus NRRL B-50147 or a mutant thereof;

the strain having the deposit accession number NRRL B-50622 or a strain having all of the identifying characteristics of Bacillus NRRL B-50622 or a mutant thereof;

the strain having the deposit accession number NRRL B-50623 or a strain having all of the identifying characteristics of Bacillus NRRL B-50623 or a mutant thereof;

the strain having the deposit accession number NRRL B-50605 or a strain having all of the identifying characteristics of Bacillus NRRL B-50605 or a mutant thereof;

the strain having the deposit accession number NRRL B-50621 or a strain having all of the identifying characteristics of Bacillus NRRL B-50621 or a mutant thereof;

the strain having the deposit accession number NRRL B-50015 or a strain having all of the identifying characteristics of Bacillus NRRL B-50015 or a mutant thereof;

the strain having the deposit accession number NRRL B-50016 or a strain having all of the identifying characteristics of Bacillus NRRL B-50016 or a mutant thereof;

the strain having the deposit accession number NRRL B-50885 or a strain having all of the identifying characteristics of Bacillus NRRL B-50885 or a mutant thereof;

the strain having the deposit accession number NRRL B-50886 or a strain having all of the identifying characteristics of Bacillus NRRL B-50886 or a mutant thereof;

the strain having the deposit accession number NRRL B-50013 or a strain having all of the identifying characteristics of Bacillus NRRL B-50013 or a mutant thereof;

the strain having the deposit accession number NRRL B-50141 or a strain having all of the identifying characteristics of Bacillus NRRL B-50141 or a mutant thereof;

the strain having the deposit accession number NRRL B-50151 or a strain having all of the identifying characteristics of Bacillus NRRL B-50151 or a mutant thereof;

the strain having the deposit accession number NRRL B-50606 or a strain having all of the identifying characteristics of Bacillus NRRL B-50606 or a mutant thereof;

the strain having the deposit accession number PTA-7543 or a strain having all of the identifying characteristics of Bacillus PTA-7543 or a mutant thereof; the strain having the deposit accession number PTA-7547 or a strain having all of the identifying characteristics of Bacillus PTA-7547 or a mutant thereof;

the strain having the deposit accession number NRRL B-50888 or a strain having all of the identifying characteristics of Bacillus NRRL B-50888 or a mutant thereof;

the strain having the deposit accession number PTA-7549 or a strain having all of the identifying characteristics of Bacillus PTA-7549 or a mutant thereof;

the strain having the deposit accession number NRRL B-50349 or a strain having all of the identifying characteristics of Bacillus NRRL B-50349 or a mutant thereof; or any combination thereof,

the strain having the deposit accession number PTA-7541 or a strain having all of the identifying characteristics of Bacillus PTA-7541 or a mutant thereof;

the strain having the deposit accession number NRRL B-59648 or a strain having all of the identifying characteristics of Bacillus NRRL B-59648 or a mutant thereof;

the strain having the deposit accession number NRRL B-59650 or a strain having all of the identifying characteristics of Bacillus NRRL B-59650 or a mutant thereof;

the strain having the deposit accession number NRRL B-59651 or a strain having all of the identifying characteristics of Bacillus NRRL B-59651 or a mutant thereof;

the strain having the deposit accession number NRRL B-676141 or a strain having all of the identifying characteristics of Bacillus NRRL B-676141 or a mutant thereof; and

the strain having the deposit accession number NRRL B-676140 or a strain having all of the identifying characteristics of Bacillus NRRL B-676140 or a mutant thereof; or

any combination thereof,

wherein the Bacillus strain improves the in vitro true digestibility and/or neutral detergent fiber digestibility of the animal feed.

3. A method for improving the digestibility of an animal feed such as a ruminant feed, comprising adding to the animal feed a Bacillus strain selected from the group consisting of: the strain having the deposit accession number ATCC 700385 or a strain having all of the identifying characteristics of Bacillus ATCC 700385 or a mutant thereof;

the strain having the deposit accession number NRRL B-50136 or a strain having all of the identifying characteristics of Bacillus NRRL B-50136 or a mutant thereof;

the strain having the deposit accession number NRRL B-50147 or a strain having all of the identifying characteristics of Bacillus NRRL B-50147 or a mutant thereof;

the strain having the deposit accession number NRRL B-50622 or a strain having all of the identifying characteristics of Bacillus NRRL B-50622 or a mutant thereof;

the strain having the deposit accession number NRRL B-50623 or a strain having all of the identifying characteristics of Bacillus NRRL B-50623 or a mutant thereof; the strain having the deposit accession number NRRL B-50605 or a strain having all of the identifying characteristics of Bacillus NRRL B-50605 or a mutant thereof;

the strain having the deposit accession number NRRL B-50621 or a strain having all of the identifying characteristics of Bacillus NRRL B-50621 or a mutant thereof;

the strain having the deposit accession number NRRL B-50015 or a strain having all of the identifying characteristics of Bacillus NRRL B-50015 or a mutant thereof;

the strain having the deposit accession number NRRL B-50016 or a strain having all of the identifying characteristics of Bacillus NRRL B-50016 or a mutant thereof;

the strain having the deposit accession number NRRL B-50885 or a strain having all of the identifying characteristics of Bacillus NRRL B-50885 or a mutant thereof;

the strain having the deposit accession number NRRL B-50886 or a strain having all of the identifying characteristics of Bacillus NRRL B-50886 or a mutant thereof;

the strain having the deposit accession number NRRL B-50013 or a strain having all of the identifying characteristics of Bacillus NRRL B-50013 or a mutant thereof;

the strain having the deposit accession number NRRL B-50141 or a strain having all of the identifying characteristics of Bacillus NRRL B-50141 or a mutant thereof;

the strain having the deposit accession number NRRL B-50151 or a strain having all of the identifying characteristics of Bacillus NRRL B-50151 or a mutant thereof;

the strain having the deposit accession number NRRL B-50606 or a strain having all of the identifying characteristics of Bacillus NRRL B-50606 or a mutant thereof;

the strain having the deposit accession number PTA-7543 or a strain having all of the identifying characteristics of Bacillus PTA-7543 or a mutant thereof;

the strain having the deposit accession number PTA-7547 or a strain having all of the identifying characteristics of Bacillus PTA-7547 or a mutant thereof;

the strain having the deposit accession number NRRL B-50888 or a strain having all of the identifying characteristics of Bacillus NRRL B-50888 or a mutant thereof;

the strain having the deposit accession number PTA-7549 or a strain having all of the identifying characteristics of Bacillus PTA-7549 or a mutant thereof;

the strain having the deposit accession number NRRL B-50349 or a strain having all of the identifying characteristics of Bacillus NRRL B-50349 or a mutant thereof;

the strain having the deposit accession number PTA-7541 or a strain having all of the identifying characteristics of Bacillus PTA-7541 or a mutant thereof;

the strain having the deposit accession number NRRL B-59648 or a strain having all of the identifying characteristics of Bacillus NRRL B-59648 or a mutant thereof;

the strain having the deposit accession number NRRL B-59650 or a strain having all of the identifying characteristics of Bacillus NRRL B-59650 or a mutant thereof; the strain having the deposit accession number NRRL B-59651 or a strain having all of the identifying characteristics of Bacillus NRRL B-59651 or a mutant thereof;

the strain having the deposit accession number NRRL B-676141 or a strain having all of the identifying characteristics of Bacillus NRRL B-676141 or a mutant thereof; and

the strain having the deposit accession number NRRL B-676140 or a strain having all of the identifying characteristics of Bacillus NRRL B-676140 or a mutant thereof; or

any combination thereof.

4. A method for improving the digestibility of an animal feed such as a ruminant feed, comprising adding to the animal feed a Bacillus strain selected from the group consisting of: the strain having the deposit accession number NRRL B-50605 or a strain having all of the identifying characteristics of Bacillus NRRL B-50605 or a mutant thereof;

the strain having the deposit accession number NRRL B-50885 or a strain having all of the identifying characteristics of Bacillus NRRL B-50885 or a mutant thereof;

the strain having the deposit accession number NRRL B-50886 or a strain having all of the identifying characteristics of Bacillus NRRL B-50886 or a mutant thereof;

the strain having the deposit accession number PTA-7541 or a strain having all of the identifying characteristics of Bacillus PTA-7541 or a mutant thereof;

the strain having the deposit accession number NRRL B-59648 or a strain having all of the identifying characteristics of Bacillus NRRL B-59648 or a mutant thereof;

the strain having the deposit accession number NRRL B-59650 or a strain having all of the identifying characteristics of Bacillus NRRL B-59650 or a mutant thereof;

the strain having the deposit accession number NRRL B-59651 or a strain having all of the identifying characteristics of Bacillus NRRL B-59651 or a mutant thereof;

the strain having the deposit accession number NRRL B-676141 or a strain having all of the identifying characteristics of Bacillus NRRL B-676141 or a mutant thereof; and

the strain having the deposit accession number NRRL B-676140 or a strain having all of the identifying characteristics of Bacillus NRRL B-676140 or a mutant thereof; or any combination thereof.

5. The method of any of items 1 or 4, wherein the animal feed is fed to a ruminant.

6. The method of any of items 1 to 5, wherein the animal feed comprises forage and optionally one or more components selected from the list consisting of:

concentrate;

one or more enzymes;

one or more additional microbes; one or more vitamins;

one or more minerals;

one or more amino acids;and

one or more other feed ingredients.

7. The method of any of items 1 to 6, wherein the ruminant feed has a 48 hour in vitro NDF Digestibility of 30-70% of NDF such as a 48 hour in vitro NDF Digestibility of NDF selected from the group consisting of from 30-40%, for example from 40-50%, such as from 50-60%, for example from 60-70%, or any combination of these intervals.

8. The method of any of items 1 to 7, wherein the animal feed is sub-optimal.

9. The method of any of items 1 to 8, wherein the method increases the milk yield and/or the meat yield of the ruminant.

10. The method of any of items 1 to 9, wherein the bacterial count of each of the bacterial strains is between 1 x10 4 and 1x10 14 CFU/kg of forage, preferably between 1 x10 6 and 1 x10 12 CFU/kg of forage, and more preferably between 1x10 7 and 1 x10 11 CFU/kg of forage and/or between 1x10 5 and 1x10 15 CFU/animal/day, preferably between 1x10 7 and 1 x10 13 CFU/animal/day, and more preferably between 1x10 8 and 1x10 12 CFU/animal/day.

1 1. A method for improving the digestibility of an animal feed such as a ruminant feed comprising:

(a) feeding an animal a feed such as a ruminant; and

(b) administering to an animal such as an ruminant a Bacillus strain selected from the group consisting of Bacillus pumilus, Bacillus amyloliquefaciens, Bacillus subtilis and Bacillus licheniformis wherein the Bacillus strain improves the in vitro true digestibility and/or neutral detergent fiber digestibility of the animal feed and wherein the Bacillus strain has enzyme activity against Casein and/or Xylan under aerobic conditions (e.g. determined as described in Example 1 to 4.

12. The method according to item 1 1 comprising:

(a) feeding an animal a feed such as a ruminant feed; and

(b) administering to an animal such as a ruminant a Bacillus strain selected from the group consisting of: the strain having the deposit accession number ATCC 700385; or a strain having all of the identifying characteristics of Bacillus ATCC 700385 or a mutant thereof;

the strain having the deposit accession number NRRL B-50136; or a strain having all of the identifying characteristics of Bacillus NRRL B-50136 or a mutant thereof;

the strain having the deposit accession number NRRL B-50147; or a strain having all of the identifying characteristics of Bacillus NRRL B-50147 or a mutant thereof;

the strain having the deposit accession number NRRL B-50622; or a strain having all of the identifying characteristics of Bacillus NRRL B-50622 or a mutant thereof;

the strain having the deposit accession number NRRL B-50623; or a strain having all of the identifying characteristics of Bacillus NRRL B-50623 or a mutant thereof;

the strain having the deposit accession number NRRL B-50605; or a strain having all of the identifying characteristics of Bacillus NRRL B-50605 or a mutant thereof;

the strain having the deposit accession number NRRL B-50621 ; or a strain having all of the identifying characteristics of Bacillus NRRL B-50621 or a mutant thereof;

the strain having the deposit accession number NRRL B-50015; or a strain having all of the identifying characteristics of Bacillus NRRL B-50015 or a mutant thereof;

the strain having the deposit accession number NRRL B-50016; or a strain having all of the identifying characteristics of Bacillus NRRL B-50016 or a mutant thereof;

the strain having the deposit accession number NRRL B-50885; or a strain having all of the identifying characteristics of Bacillus NRRL B-50885 or a mutant thereof;

the strain having the deposit accession number NRRL B-50886; or a strain having all of the identifying characteristics of Bacillus NRRL B-50886 or a mutant thereof;

the strain having the deposit accession number NRRL B-50013; or a strain having all of the identifying characteristics of Bacillus NRRL B-50013 or a mutant thereof; the strain having the deposit accession number NRRL B-50141 ; or a strain having all of the identifying characteristics of Bacillus NRRL B-50141 or a mutant thereof;

the strain having the deposit accession number NRRL B-50151 ; or a strain having all of the identifying characteristics of Bacillus NRRL B-50151 or a mutant thereof;

the strain having the deposit accession number NRRL B-50606; or a strain having all of the identifying characteristics of Bacillus NRRL B-50606 or a mutant thereof;

the strain having the deposit accession number PTA-7543; or a strain having all of the identifying characteristics of Bacillus PTA-7543 or a mutant thereof;

the strain having the deposit accession number PTA-7547; or a strain having all of the identifying characteristics of Bacillus PTA-7547 or a mutant thereof;

the strain having the deposit accession number NRRL B-50888; or a strain having all of the identifying characteristics of Bacillus NRRL B-50888 or a mutant thereof;

the strain having the deposit accession number PTA-7549; or a strain having all of the identifying characteristics of Bacillus PTA-7549 or a mutant thereof;

the strain having the deposit accession number NRRL B-50349; or a strain having all of the identifying characteristics of Bacillus NRRL B-50349 or a mutant thereof,

the strain having the deposit accession number PTA-7541 or a strain having all of the identifying characteristics of Bacillus PTA-7541 or a mutant thereof;

the strain having the deposit accession number NRRL B-59648 or a strain having all of the identifying characteristics of Bacillus NRRL B-59648 or a mutant thereof;

the strain having the deposit accession number NRRL B-59650 or a strain having all of the identifying characteristics of Bacillus NRRL B-59650 or a mutant thereof;

the strain having the deposit accession number NRRL B-59651 or a strain having all of the identifying characteristics of Bacillus NRRL B-59651 or a mutant thereof;

the strain having the deposit accession number NRRL B-676141 or a strain having all of the identifying characteristics of Bacillus NRRL B-676141 or a mutant thereof; and the strain having the deposit accession number NRRL B-676140 or a strain having all of the identifying characteristics of Bacillus NRRL B-676140 or a mutant thereof; or any combination thereof,

wherein the Bacillus strain improves the in vitro true digestibility and/or neutral detergent fiber digestibility of the animal feed, and wherein step (a) occurs before, after, or simultaneously with step (b).

13. A method for improving the digestibility of an animal feed such as a ruminant feed comprising:

(a) feeding an animal a feed such as a ruminant feed; and

(b) administering to an animal such as a ruminant a Bacillus strain selected from the group consisting of:

the strain having the deposit accession number ATCC 700385; or a strain having all of the identifying characteristics of Bacillus ATCC 700385 or a mutant thereof;

the strain having the deposit accession number NRRL B-50136; or a strain having all of the identifying characteristics of Bacillus NRRL B-50136 or a mutant thereof;

the strain having the deposit accession number NRRL B-50147; or a strain having all of the identifying characteristics of Bacillus NRRL B-50147 or a mutant thereof;

the strain having the deposit accession number NRRL B-50622; or a strain having all of the identifying characteristics of Bacillus NRRL B-50622 or a mutant thereof;

the strain having the deposit accession number NRRL B-50623; or a strain having all of the identifying characteristics of Bacillus NRRL B-50623 or a mutant thereof;

the strain having the deposit accession number NRRL B-50605; or a strain having all of the identifying characteristics of Bacillus NRRL B-50605 or a mutant thereof;

the strain having the deposit accession number NRRL B-50621 ; or a strain having all of the identifying characteristics of Bacillus NRRL B-50621 or a mutant thereof;

the strain having the deposit accession number NRRL B-50015; or a strain having all of the identifying characteristics of Bacillus NRRL B-50015 or a mutant thereof; the strain having the deposit accession number NRRL B-50016; or a strain having all of the identifying characteristics of Bacillus NRRL B-50016 or a mutant thereof;

the strain having the deposit accession number NRRL B-50885; or a strain having all of the identifying characteristics of Bacillus NRRL B-50885 or a mutant thereof;

the strain having the deposit accession number NRRL B-50886; or a strain having all of the identifying characteristics of Bacillus NRRL B-50886 or a mutant thereof;

the strain having the deposit accession number NRRL B-50013; or a strain having all of the identifying characteristics of Bacillus NRRL B-50013 or a mutant thereof;

the strain having the deposit accession number NRRL B-50141 ; or a strain having all of the identifying characteristics of Bacillus NRRL B-50141 or a mutant thereof;

the strain having the deposit accession number NRRL B-50151 ; or a strain having all of the identifying characteristics of Bacillus NRRL B-50151 or a mutant thereof;

the strain having the deposit accession number NRRL B-50606; or a strain having all of the identifying characteristics of Bacillus NRRL B-50606 or a mutant thereof;

the strain having the deposit accession number PTA-7543; or a strain having all of the identifying characteristics of Bacillus PTA-7543 or a mutant thereof;

the strain having the deposit accession number PTA-7547; or a strain having all of the identifying characteristics of Bacillus PTA-7547 or a mutant thereof;

the strain having the deposit accession number NRRL B-50888; or a strain having all of the identifying characteristics of Bacillus NRRL B-50888 or a mutant thereof;

the strain having the deposit accession number PTA-7549; or a strain having all of the identifying characteristics of Bacillus PTA-7549 or a mutant thereof;

the strain having the deposit accession number NRRL B-50349; or a strain having all of the identifying characteristics of Bacillus NRRL B-50349 or a mutant thereof,

the strain having the deposit accession number PTA-7541 or a strain having all of the identifying characteristics of Bacillus PTA-7541 or a mutant thereof; the strain having the deposit accession number NRRL B-59648 or a strain having all of the identifying characteristics of Bacillus NRRL B-59648 or a mutant thereof;

the strain having the deposit accession number NRRL B-59650 or a strain having all of the identifying characteristics of Bacillus NRRL B-59650 or a mutant thereof;

the strain having the deposit accession number NRRL B-59651 or a strain having all of the identifying characteristics of Bacillus NRRL B-59651 or a mutant thereof;

the strain having the deposit accession number NRRL B-676141 or a strain having all of the identifying characteristics of Bacillus NRRL B-676141 or a mutant thereof; and

the strain having the deposit accession number NRRL B-676140 or a strain having all of the identifying characteristics of Bacillus NRRL B-676140 or a mutant thereof; or

any combination thereof, and

wherein step (a) occurs before, after, or simultaneously with step (b).

14. The method of any of items 1 1 to 13, wherein the animal feed is fed to a ruminant.

15. The method of any of items 1 1 to 14, wherein the animal feed comprises forage and optionally one or more components selected from the list consisting of:

concentrate;

one or more enzymes;

one or more additional microbes;

one or more vitamins;

one or more minerals;

one or more amino acids;and

one or more other feed ingredients.

16. The method of any of items 1 1 to 15, wherein the ruminant feed has a 48 hour in vitro NDF Digestibility of 30-70% of NDF such as a 48 hour in vitro NDF Digestibility of NDF selected from the group consisting of from 30-40%, for example from 40-50%, such as from 50-60%, for example from 60-70%, or any combination of these intervals.

17. The method of any of items 1 1 to 16, wherein the animal feed is sub-optimal. 18. The method of any of items 1 1 to 17, wherein the method increases the milk yield and/or the meat yield of the ruminant.

19. The method of any of items 1 1 to 18, wherein the bacterial count of each of the bacterial strains is between 1x10 4 and 1 x10 14 CFU/kg of forage, preferably between 1 x10 6 and 1 x10 12 CFU/kg of forage, and more preferably between 1 x10 7 and 1 x10 11 CFU/kg of forage and/or between 1x10 5 and 1 x10 15 CFU/animal/day, preferably between 1 x10 7 and 1x10 13 CFU/animal/day, and more preferably between 1x10 8 and 1x10 12 CFU/animal/day. 20. An animal feed composition such as a ruminant feed composition comprising forage and a Bacillus strain selected from the group consisting of:

the strain having the deposit accession number NRRL B-50885; or a strain having all of the identifying characteristics of NRRL B-50885 or a mutant thereof;

the strain having the deposit accession number NRRL B-50886; or a strain having all of the identifying characteristics of NRRL B-50886 or a mutant thereof;

the strain having the deposit accession number NRRL B-50888 or a strain having all of the identifying characteristics of NRRL B-50888 or a mutant thereof;

the strain having the deposit accession number PTA-7541 or a strain having all of the identifying characteristics of Bacillus PTA-7541 or a mutant thereof;

the strain having the deposit accession number NRRL B-59648 or a strain having all of the identifying characteristics of Bacillus NRRL B-59648 or a mutant thereof;

the strain having the deposit accession number NRRL B-59650 or a strain having all of the identifying characteristics of Bacillus NRRL B-59650 or a mutant thereof;

the strain having the deposit accession number NRRL B-59651 or a strain having all of the identifying characteristics of Bacillus NRRL B-59651 or a mutant thereof;

the strain having the deposit accession number NRRL B-676141 or a strain having all of the identifying characteristics of Bacillus NRRL B-676141 or a mutant thereof; and

the strain having the deposit accession number NRRL B-676140 or a strain having all of the identifying characteristics of Bacillus NRRL B-676140 or a mutant thereof; or any combination thereof.

21. The animal feed composition of item 20 further comprising a bacterium from one or more of the following genera: Lactobacillus, Lactococcus, Streptococcus, Bacillus, Pediococcus, Enterococcus, Leuconostoc, Carnobacterium, Propionibacterium, Bifidobacterium, Clostridium and Megasphaera or any combination thereof. 22. The animal feed composition of any of items 20 to 21 further comprising a Bacillus strain selected from the group consisting of:

the si rain having the depos accession number ATCC 700385;

the si rain having the depos accession number NRRL B-50136;

the si rain having the depos accession number NRRL B-50622;

the si rain having the depos accession number NRRL B-50623;

the si rain having the depos accession number NRRL B-50605;

the si rain having the depos accession number NRRL B-50621 ;

the si rain having the depos accession number NRRL B-50015;

the si rain having the depos accession number NRRL B-50016;

the si rain having the depos accession number NRRL B-50013;

the si rain having the depos accession number NRRL B-50141 ;

the si rain having the depos accession number NRRL B-50151 ;

the si rain having the depos accession number NRRL B-50606;

the si rain having the depos accession number PTA-7543;

the si rain having the depos accession number PTA-7547;

the si rain having the depos accession number PTA-7549; and

the si rain having the depos accession number NRRL B-50349,

the si rain having the depos accession number PTA-7541 ;

the si rain having the depos accession number NRRL B-59648;

the si rain having the depos accession number NRRL B-59650;

the si rain having the depos accession number NRRL B-59651 ;

the si rain having the depos accession number NRRL B-676141 ; and

the si rain having the depos accession number NRRL B-676140; or

any combination thereof.

23. The animal feed composition of any of items 20 to 22, wherein one or more of the bacterial strains are in spore form. 24. The animal feed composition of any of items 20 to 23, wherein the bacterial count of each of the bacterial strains is between 1x10 4 and 1x10 14 CFU/kg of forage, preferably between 1 x10 6 and 1 x10 12 CFU/kg of forage, and more preferably between 1 x10 7 and 1 x10 11 CFU/kg of forage and/or between 1x10 5 and 1 x10 15 CFU/animal/day, preferably between 1x10 7 and 1 x10 13 CFU/animal/day, and more preferably between 1 x10 8 and 1x10 12 CFU/animal/day. 25. The animal feed composition of any of items 20 to 24, wherein the animal feed comprises forage and optionally one or more components selected from the list consisting of: concentrate;

one or more enzymes;

one or more additional microbes;

one or more vitamins;

one or more minerals;

one or more amino acids;and

one or more other feed ingredients.

26. A Bacillus strain selected from the group consisting of

the strain having the deposit accession number PTA-7541 or a strain having all of the identifying characteristics of Bacillus PTA-7541 or a mutant thereof;

the strain having the deposit accession number NRRL B-676141 or a strain having all of the identifying characteristics of Bacillus NRRL B-676141 or a mutant thereof; and

the strain having the deposit accession number NRRL B-676140 or a strain having all of the identifying characteristics of Bacillus NRRL B-676140 or a mutant thereof.

Examples

Assays

In vitro True Digestibility and Neutral Detergent Fiber Digestibility

Following addition of Bacillus strains, in vitro true digestibility (IVTD) and Neutral Detergent Fiber Digestibility (NDFD) studies were conducted by Dairy One Forage Laboratory (Ithaca, NY, USA). The substrate was removed from each bag, dried for 4 hours at 60°C, and then ground through a 1 mm UDY Cyclone Mill (UDY Corp., Fort Collins, CO, USA). Totally 250 mg of dried, milled substrate were incubated in Van Soest buffer (Goering and Van Soest, 1970, Forage fiber analysis (apparatus, reagents, procedures and some applications), Agricultural Handbook No. 379 ARS-USDA, Washington, DC with rumen fluid from high producing dairy cows consuming a typical Total Mixed Rations (TMR) diet. The incubations were performed at 39°C for 24 hours in ANOKM® F57 filter bags (ANOKM Technology, Macedon, NY, USA). After incubation, samples of the undigested fibrous residue were determined using an NDF procedure (ANKOM A200 Filter Bag Technique (FBT), ANKOM Application Note 01/02 "Method for Determining Neutral Detergent Fiber (aNDF)". Solutions are described in Journal of Dairy Science 74: 3583-3597 (1991 )). The IVTD and NDFD were determined by the undigested fibrous residue remaining after digestion. An increase in digestibility was determined by comparison of the average % of digested material for the untreated control compared to the average percent of digested material for the microbial treatment samples.

Example 1 : Determination of IVTD and NDFD on Corn Silage

A total of 50 gm of mature corn silage (conventional hybrid, processed, chopped at

7/8" in length with 34% dry mater) was inoculated separately with twenty different Bacillus strains at a rate of approximately 1 x10 6 CFU/gm corn silage in 1 gallon vacuum bags. The Bacillus strains are given in tables 1 -3 below.

The microbes were thoroughly mixed with the corn silage. A vacuum was applied and the bags were sealed using a commercially available vacuum system creating an anaerobic environment. IVTD and NDFD were determined as described in the "In vitro True Digestibility and Neutral Detergent Fiber Digestibility" assay above and the results are presented in Tables 1-3 and Figures 1 -3.

Table 1 : IVT and NDFD of 7 Bacillus strains on Corn Silage

NRRL B-50141 79.8 51.3

NRRL B-50151 79.8 51.3

Control 80.0 50.3

Table 3: IVT and NDFD of 6 Bacillus strains on Corn Silage

The results from Tables 1 , 2 and 3 show an increase in the percentage of neutral detergent fiber digestibility for Bacilli NRRL B-50147, NRRL B-50622, NRRL B-50621 , NRRL B-50015, NRRL B-50151 , PTA-7543, PTA-7547, NRRL B-50888, PTA-7549 and NRRL B- 50349.

Furthermore, the results from Tables 1 , 2 and 3 show an increased mature corn silage in vitro true digestibility for Bacilli ATCC 700385, NRRL B-50136, NRRL B-50147, NRRL B-50622, NRRL B-50623, NRRL B-50605, NRRL B-50621 , PTA-7543, PTA-7547, NRRL B-50888, PTA-7549 and NRRL B-50349.

Example 2: Determination of IVTD and NDFD on Timothy Hay

A total of 50 gm of Timothy hay (conventional hybrid, processed, chopped at 7/8" in length with 34% dry mater) was inoculated separately with twenty different Bacillus strains at a rate of approximately 1 x10 6 CFU/gm Timothy hay in 1 gallon vacuum bags. The Bacillus strains are given in tables 4-6 below.

The microbes were thoroughly mixed with the Timothy hay. A vacuum was applied and the bags were sealed using a commercially available vacuum system creating an anaerobic environment. IVTD and NDFD were determined as described in the "In vitro True Digestibility and Neutral Detergent Fiber Digestibility" assay above and the results are presented in Tables 4-6 and Figures 4-6. Table 4: IVT and NDFD of 7 Bacillus strains on Timothy hay

Table 5: IVT and NDFD of 7 Bacillus strains on Timothy hay

Table 6: IVT and NDFD of 7 Bacillus strains on Timothy hay NRRL B-50888 74.5 58.0

ATCC PTA-7549 73.8 53.8

NRRL B-50349 69.0 51 .5

Control 67.0 47.8

The results from Tables 4, 5 and 6 show an increase in the percentage of neutral detergent fiber digestibility on Timothy hay for Bacilli ATCC 700385, NRRL B-50147, NRRL B-50622, NRRL B-50623, NRRL B-50605, NRRL B-50621 , NRRL B-50015, NRRL B-50885, NRRL B-50886, NRRL B-50013, NRRL B-50141 , NRRL B-50151 , NRRL B-50606, PTA- 7543, PTA-7547, NRRL B-50888, PTA-7549 and NRRL B-50349.

Furthermore, the results from tables 4, 5 and 6 show an increase in vitro true digestibility on Timothy hay for Bacilli ATCC 700385, NRRL B-50136, NRRL B-50147, NRRL B-50622, NRRL B-50623, NRRL B-50605, NRRL B-50621 , NRRL B-50015, NRRL B-50016, NRRL B-50885, NRRL B-50886, NRRL B-50013, NRRL B-50141 , NRRL B-50151 , NRRL B- 50606, PTA-7543, PTA-7547, NRRL B-50888, PTA-7549 and NRRL B-50349.

Example 3: Results for AZCL substrate testing of enzyme activities

Commercially available AZCL substrates were incorporated into agar plates to monitor enzyme activity based on liberating a soluble blue dye fragment. Culture supernatants from an overnight culture were spotted on AZCL substrate plates and incubated for 30 °C for 24 h (Aerobic incubation) and 48 h (Anaerobic incubation). Observation of the liberated soluble blue dye as a result of enzyme hydrolysis served as evidence of enzyme activity present for a particular substrate. All samples and controls were run in duplicate wells. Non-inoculated wells were included as negative controls (n=6 wells), and enzyme controls (n=2 wells) were included as positive controls. The results are shown in Table 7 and Table 8 herein below. Furthermore the results from Table 7 and Table 8, show strain to strain variation in enzyme activity both aerobically and anaerobically, with consistent positive activity on casein substrate under aerobic growth conditions.

Table 7: AZCL Activity - Aerobic

NRRL B-50623 + + + + - +

NRRL B-50605 + + + + + +

NRRL B-50621 + + - + - -

NRRL B-50015 + + - + - -

NRRL B-50016 - - + + - +

NRRL B-50885 - - - + - +

NRRL B-50886 - - - + - +

NRRL B-50013 + + + + + +

NRRL B-50141 + + + + + +

NRRL B-50151 + + + + + +

NRRL B-50606 + + + + - +

PTA-75 ' 43 + + + + - +

PTA-7547 + + + + + +

NRRL B-50888 + + + + + +

PTA-7549 + + + + - +

NRRL B-50349 + + + + + +

PTA-7541 + + + + - +

NRRL B-59648 + + + + + +

NRRL B-59650 + + + + - +

NRRL B-59651 - - + + - +

NRRL B-676141 - - + + - +

NRRL B-676140 + + + + + +

Table 8: ZCL Activity - Anaerobic

All samples were ran in duplicate. "+" indicates both wells showed activity, "-" indicates neither well showed activity, and "+/-" or "-/+" indicated one of two wells had activity. A entry of "weak" indicates that the color of dye liberated was low, but observable. Example 4: Determination of NVTD and ADFD

In vitro Semi-Continuous Neutral Detergent Fiber Digestibility (NDFD) and Acid Detergent Fiber Digestibility (ADFD) (Knap et al., 2010, Bacillus licheniformis Prevents Necrotic Enteritis in Broiler Chickens, Avian Diseases 54(2): 931-935)

Following addition of Bacillus strains, in vitro Neutral Detergent Fiber Digestibility

(NDFD) and Acid Detergent Fiber Digestibility (ADFD) studies were conducted by West Virginia University (Morgantown, WV, USA). Diets were fed as a total mixed ration with all of the ingredients ground to pass through a 4 mm screen attached to a Wiley Mill (Thomas Scientific, Swedesboro, NJ, USA). Ruminal fluid was collected 3 hours after feeding and 1 L of fluid was added to each of 12 dual-flow continuous culture apparatuses (Stern and Hoover, 1990 [The dual flow continuous culture system. Pages 17-32 in Proc. Continuous Culture Fermenters: Frustration or Fermentation. Northwest ADSA-ASAS Regional Meeting, Chazy, NY. Am. Dairy Sci. Assoc., Savoy, IL.]) along with 200 mL of buffer (Weller and Pilgrim, 1974, omitting urea [Passage of protozoa and volatile fatty acids from the rumen of sheep and from a continuous in vitro fermentation system. Br. J. Nutr. 32: 341 -352]). 25 g of dried, milled substrate was fed 4 times per day at 6 hour intervals for a total of 100 g dry matter fed per day. The liquid dilution rate was 13% per hour with a solids retention time of 22 hours.

The ADFD and NDFD were determined by the undigested fibrous residue remaining after digestion. Effluent dry matter was determined by centrifuging a weighed sample and drying the precipitated residue. The ADF and NDF concentrations in the feed and effluent were determined with the method of Van Soest, 1991 [Methods for dietary fiber, neutral detergent fiber, and nonstarch polysaccharides in relation to animal nutrition. J. Dairy Sci. 74: 3583-3597]. An increase in digestibility was determined by comparison of the average % of digested material for the untreated control compared to the average percent of digested material for the microbial treatment samples.

Diets used consisted of a 20/20/20% blend of alfalfa balage, corn silage, and grass hay or a 12.5/35/12.5% blend of each of the respective components. The percent of each ingredient and component analysis for each of three trials are outlined in Table 9. A total of 4.5 kg of the mixed ration diet was inoculated separately with nine different Bacillus strains at a rate of approximately 4x10 4 CFU/g dry matter mixed ration. The Bacillus strains are given in Table 10 below. The microbes were thoroughly mixed with the forages and all ingredients ground together through a 4 mm screen. 25 g of dried, milled substrate was fed 4 times per day at 6 hour intervals for a total of 100 g dry matter fed per day. Feed was added to a dual- flow continuous culture apparatus (Stern and Hoover, 1990 [The dual flow continuous culture system. Pages 17-32 in Proc. Continuous Culture Fermenters: Frustration or Fermentation. Northwest ADSA-ASAS Regional Meeting, Chazy, NY. Am. Dairy Sci. Assoc., Savoy, IL.]). The liquid dilution rate was 13% per hour with a solids retention time of 22 hours.

Table 9: Components analysis of mixed ration diets used

The NDFD and ADFD were determined by the undigested fibrous residue remaining after digestion. An increase in digestibility was determined by comparison of the average % of digested material for the untreated control compared to the average percent of digested material for the microbial treatment samples.

Table 10: NDFD and ADFD of 9 Bacillus strains on mixed ration diet

* Represents the average from CCIV#1 and 2

* * represents the average from CCIV#1 , 2 and 3

The results from table 2 shows an increase in the percentage of NDFD compared to control for Bacilli NRRL B-50885, NRRL B-50886, NRRL B-50605, PTA-7541 , NRRL B- 59648, NRRL B-59650, NRRL B-59651 , and NRRL B-676141 .

Furthermore, the results from tables 2 show an increase in the percentage of ADFD compared to control for Bacilli NRRL B-50885, NRRL B-50605, PTA-7541 , NRRL B-676141 , and NRRL B-676140.

Deposit of Biological Material

The following biological materials were deposited under the terms of the Budapest

Treaty at the American Type Culture Collection (ATCC), 10801 University Blvd., Manassas, VA 20108, USA, and the Microbial Genomics and Bioprocessing Research Unit (NRRL) National Center for Agricultural Utilization Research 1815 N. University Street, Peoria, IL 61604, USA and given the following accession numbers: Table 11: Deposit of Biological Material

NRRL B-50885 was isolated from hydrocarbon contaminated soil, southern Virginia, USA; NRRL B-50886 was isolated from hydrocarbon contaminated soil, southern Virginia, USA; ; NRRL-B-59650 was isolated from a human axilla cup-scrub sample from a subject in southern Virginia, USA; NRRL-B-59651 was isolated from a human axilla cup-scrub sample from a subject in southern Virginia, USA; and NRRL B-50888 was isolated from a tire Disposal site, Roanoke, Virginia, USA.

The strains have been deposited under conditions that assure that access to the culture will be available during the pendency of this patent application to one determined by foreign patent laws to be entitled thereto. The deposits represent a substantially pure culture of the deposited strain. The deposits are available as required by foreign patent laws in countries wherein counterparts of the subject application or its progeny are filed. However, it should be understood that the availability of a deposit does not constitute a license to practice the subject invention in derogation of patent rights granted by governmental action.