Microbial compositions for inhibiting fusobacterium or trueperella

Abstract

The invention relates to direct-fed microbials for use in controlling diseases in animals caused by Fusobacterium or Trueperella species. More particularly, the invention relates to isolated Bacillus strains 86, 300, 101, 235, 177, and 102, and strains having all of the identifying characteristics of these strains, for uses comprising the above-mentioned uses.

Description

MICROBIAL COMPOSITIONS FOR INHIBITING FUSOBACTERIUM OR TRUEPERELLACROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit under 35 USC § 119(e) of U.S. Provisional Application Serial No. 63 / 461,720, filed on April 25, 2023, the entire disclosure of which is incorporated herein by reference.REFERENCE TO AN ELECTRONICALLY SUBMITTED SEQUENCE LISTING

[0002] The contents of the electronic sequence listing (file name: 60421409182.xml; size: 4 kilobytes, created on April 22, 2024) is herein incorporated by reference in its entirety.FIELD OF THE DISCLOSURE

[0003] The invention relates to direct-fed microhials for use in controlling diseases in animals caused by Fusobacterium or Trueperella species. More particularly, the invention relates to isolated Bacillus strains 86, 300, 101, 235, 177, and 102, and strains having all of the identifying characteristics of these strains, for uses comprising the above-mentioned uses.BACKGROUND AND SUMMARY OF THE INVENTION

[0004] The present invention relates to direct-fed microbial (DFM) compositions.Liver abscesses can be problematic for many ruminants but are typically associated with feedlot cattle where they are the most common cause of liver condemnation, with incidence typically ranging from 10-20% of feedlot animals. Reduced production efficiency is also observed in cattle with liver abscesses. Together the liver condemnation, carcass quality impact and lowered production efficiency have a substantial impact on profitability for producers.

[0005] Liver abscesses are associated with acidosis and rumenitis, in which rapid fermentation of dietary carbohydrate in the rumen increases acidity and causes areas of necrosis or lesions on the interior rumen wall. Pathogenic and opportunistic bacteria colonize the compromised rumen tissue and are transported to the liver through the hepatic portal vein where they establish and cause the development of necrobacillosis and abscess. Bacterial translocation from other portions of the gut to the liver may also contribute to liver abscess development.

[0006] Fusobacterium necrophorum, a Gram-negative anaerobe, has been identified as the primary microbial agent in the development of bovine liver abscesses and has been implicated as the primary cause of necrotic laryngitis (calf diphtheria) and of localized or mixedinfections including foot rot, lameness, necrobacillosis, and abscesses in sheep, deer, other ruminants, and swine. Fusobacterium varium, though less frequently reported, can also be found in liver abscesses and can be highly resistant to antibiotics.

[0007] Trueperella pyogenes (formerly Actinomyces pyogenes) is a Gram-positive, facultative anaerobe that is found in the rumen but also in the urogenital, gastrointestinal, and upper respiratory tracts of mammals and is involved in multiple opportunistic infections including abscesses, mastitis, metritis, pyometra, and pneumonia. Fusobacterium necrophorum and Trueperella pyogenes are often found concurrently in liver abscesses. Therefore, technologies to help manage or reduce levels of Fusobacterium necrophorum, Trueperella pyogenes and other opportunistic microorganisms within the rumen and gastrointestinal tract are beneficial for reducing translocation to the liver and reducing frequency and severity of liver abscesses and associated diseases.

[0008] Applicants have developed a direct-fed microbial composition that reduces the growth of Fusobacterium necrophorum and Trueperella pyogenes associated with liver abscess and other necrobacillosis. The direct-fed microbial compositions described herein offer a commercial benefit by providing all of the above-mentioned properties, or a combination thereof, in a single direct-fed microbial composition.

[0009] Tn various embodiments, the compositions for use in the methods described herein can be a commercial package, a feed additive for an animal feed composition, an additive for the drinking water of an animal, or an animal feed composition (e.g., a complete feed), each comprising one or more of the isolated strains comprising Bacillus strain 86 (NRRL No. B-50944), Bacillus strain 300 (NRRL No. B-50943), Bacillus strain 101 (NRRL No. B- 67218), Bacillus strain 235 (NRRL No. B-67219), Bacillus strain 177 (NRRL No. B-67275), and Bacillus strain 102 (NRRL No. B-67276), or strains having all of the identifying characteristics of these strains.

[0010] In another embodiment, a method is provided of treating an animal wherein the animal has a disease caused by a Fusobacterium or Trueperella species. The method comprises the step of administering to the animal a feed composition or drinking water comprising an effective amount of an additive comprising an isolated Bacillus strain selected from the group consisting of Bacillus strain 86 (NRRL No. B-50944), Bacillus strain 300 (NRRL No. B- 50943), Bacillus strain 101 (NRRL No. B-67218), Bacillus strain 235 (NRRL No. B-67219), Bacillus strain 177 (NRRL No. B-67275), Bacillus strain 102 (NRRL No. B-67276), a strain having all of the identifying characteristics of Bacillus strain 86 (NRRL No. B-50944), a strain having all of the identifying characteristics of Bacillus strain 300 (NRRL No. B-50943), a strain having all of the identifying characteristics of Bacillus strain 101 (NRRL No. B-67218), a strainhaving all of the identifying characteristics of Bacillus strain 235 (NRRL No. B-67219), a strain having all of the identifying characteristics of Bacillus strain 177 (NRRL No. B-67275), a strain having all of the identifying characteristics of Bacillus strain 102 (NRRL No. B-67276), and combinations thereof, wherein the Bacillus strain improves the disease caused by the Fusobacterium or Trueperella species.

[0011] In yet another embodiment, a method of treating an animal is provided wherein the animal has a disease selected from the group consisting of liver abscess, necrotic laryngitis, calf diphtheria, mastitis, pyometra, and bull nose disease. The method comprises the step of administering to the animal a feed composition or drinking water comprising an effective amount of an additive comprising an isolated Bacillus strain selected from the group consisting of Bacillus strain 86 (NRRL No. B-50944), Bacillus strain 300 (NRRL No. B-50943), Bacillus strain 101 (NRRL No. B-67218), Bacillus strain 235 (NRRL No. B-67219), Bacillus strain 177 (NRRL No. B-67275), Bacillus strain 102 (NRRL No. B-67276), a strain having all of the identifying characteristics of Bacillus strain 86 (NRRL No. B-50944), a strain having all of the identifying characteristics of Bacillus strain 300 (NRRL No. B-50943), a strain having all of the identifying characteristics of Bacillus strain 101 (NRRL No. B-67218), a strain having all of the identifying characteristics of Bacillus strain 235 (NRRL No. B-67219), a strain having all of the identifying characteristics of Bacillus strain 177 (NRRL No. B-67275), a strain having all of the identifying characteristics of Bacillus strain 102 (NRRL No. B-67276), and combinations thereof, wherein the Bacillus strain improves the disease selected from the group consisting of liver abscess, necrotic laryngitis, calf diphtheria, mastitis, pyometra, and bull nose disease.

[0012] The following clauses, and combinations thereof, provide various additional illustrative aspects of the invention described herein. The various embodiments described in any other section of this patent application, including the section titled “DETAILED DESCRIPTION OF ILLUSTRATIVE EMBODIMENTS” and the EXAMPLES are applicable to any of the following embodiments of the invention described in the numbered clauses below.

[0013] 1. A method of treating an animal wherein the animal has a disease caused by a Fusobacterium or Trueperella species, the method comprising the step of administering to the animal a feed composition or drinking water comprising an effective amount of an additive comprising an isolated Bacillus strain selected from the group consisting of Bacillus strain 86 (NRRL No. B-50944), Bacillus strain 300 (NRRL No. B-50943), Bacillus strain 101 (NRRL No. B-67218), Bacillus strain 235 (NRRL No. B-67219), Bacillus strain 177 (NRRL No. B- 67275), Bacillus strain 102 (NRRL No. B-67276), a strain having all of the identifying characteristics of Bacillus strain 86 (NRRL No. B-50944), a strain having all of the identifying characteristics of Bacillus strain 300 (NRRL No. B-50943), a strain having all of the identifyingcharacteristics of Bacillus strain 101 (NRRL No. B-67218), a strain having all of the identifying characteristics of Bacillus strain 235 (NRRL No. B-67219), a strain having all of the identifying characteristics of Bacillus strain 177 (NRRL No. B-67275), a strain having all of the identifying characteristics of Bacillus strain 102 (NRRL No. B-67276), and combinations thereof, wherein the Bacillus strain improves the disease caused by the Fusobacterium or Trueperella species.

[0014] 2. The method of clause 1 wherein the animal is selected from the group consisting of a poultry species, a porcine species, a bovine species, an ovine species, a cervid species, an equine species, and a companion animal.

[0015] 3. The method of clause 1 or 2 wherein the animal is a dairy animal.

[0016] 4. The method of clause 3 wherein the dairy animal is selected from the group consisting of a cow, a goat, a sheep, and a buffalo.

[0017] 5. The method of clause 1 or 2 wherein the animal is a deer.

[0018] 6. The method of any one of clauses 1 to 5 wherein the Bacillus strain produces an enzyme selected from the group consisting of an a-galactosidase, a protease, a lipase, an amylase, a xylanase, a cellulase, and combinations thereof.

[0019] 7. The method of any one of clauses 1 to 6 further comprising the step of administering to the animal another bacterial strain selected from the group consisting of another Bacillus strain, a lactic acid bacterial strain, and combinations thereof.

[0020] 8. The method of any one of clauses 1 to 7 wherein Bacillus strain 86(NRRL No. B-50944), Bacillus strain 300 (NRRL No. B-50943), Bacillus strain 101 (NRRL No. B-67218), Bacillus strain 235 (NRRL No. B-67219), Bacillus strain 177 (NRRL No. B- 67275), and Bacillus strain 102 (NRRL No. B-67276) are all administered to the animal.

[0021] 9. The method of any one of clauses 1 to 7 wherein the strain administered is Bacillus strain 101 (NRRL No. B-67218).

[0022] 10. The method of any one of clauses 1 to 7 wherein the strain administered is Bacillus strain 235 (NRRL No. B-67219).

[0023] 11. The method of any one of clauses 1 to 7 wherein the strain administered is Bacillus strain 86 (NRRL No. B-50944).

[0024] 12. The method of any one of clauses 1 to 7 wherein the strain administered is Bacillus strain 300 (NRRL No. B-50943).

[0025] 13. The method of any one of clauses 1 to 7 wherein the strain administered is Bacillus strain 177 (NRRL No. B-67275).

[0026] 14. The method of any one of clauses 1 to 7 wherein the strain administered is Bacillus strain 102 (NRRL No. B-67276).

[0027] 15. The method of any one of clauses 1 to 7 wherein the strains administered are Bacillus strains 102 (NRRL No. B-67276) and 177 (NRRL No. B-67275).

[0028] 16. The method of any one of clauses 1 to 15 wherein the disease is caused by a Fusobacterium species.

[0029] 17. The method of clause 16 wherein the Fusobacterium species is selected from the group consisting of Fusobacterium varium, Fusobacterium necrophorum, and combinations thereof.

[0030] 18. The method of clause 16 wherein the Fusobacterium species is aFusoobacterium subspecies selected from the group consisting of Fusobacterium necrophorum, Fusobacterium necrophorum funduliforme, and combinations thereof.

[0031] 19. The method of any one of clauses 1 to 15 wherein the disease is caused by a Trueperella species.

[0032] 20. The method of clause 19 wherein the Trueperella species is Trueperella pyogenes.

[0033] 21. The method of any one of clauses 1 to 20 wherein the disease is selected from the group consisting of liver abscess, necrobacillosis, necrotic laryngitis, lameness, mastitis, metritis, pyometra, pneumonia, and combinations thereof.

[0034] 22. The method of clause 21 wherein the necrobacillosis is selected from bull nose disease and foot rot.

[0035] 23. The method of clause 21 wherein the necrotic laryngitis is calf diphtheria.

[0036] 24. The method of any one of clauses 1 to 23 wherein the improvement in the disease caused by the Fusobacterium or Trueperella species results in an improvement selected from the group consisting of an improvement in the health of the animal, an improvement in carcass quality, an improvement in production efficiency, an improvement in condemnation rate, an improvement in feed efficiency, and combinations thereof.

[0037] 25. The method of any one of clauses 1 to 24 wherein each of the Bacillus strains is administered in the feed composition at a dose of about 1.0 x 103CFU / pound of the feed composition to about 5.0 x 1012CFU / pound of the feed composition.

[0038] 26. The method of any one of clauses 1 to 24 wherein each of the Bacillus strains is administered in the feed composition at a dose of about 0.5 x 107CFU / pound of the feed composition to about 5.0 x 107CFU / pound of the feed composition.

[0039] 27. The method of any one of clauses 1 to 24 wherein each of the Bacillus strains is administered in the feed composition at a dose greater than about 5.0 x 106CFU / pound of the feed composition.

[0040] 28. The method of any one of clauses 1 to 27 further comprising the step of administering an antibiotic to the animal.

[0041] 29. The method of any one of clauses 1 to 28 further comprising the step of administering to the animal an enzyme selected from the group consisting of a galactosidase, a protease, a lipase, an amylase, a hemicellulase, an arabinoxylanase, a xylanase, a cellulase, an NSPase, a phytase, and combinations thereof.

[0042] 30. The method of clause 29 wherein the enzyme is an NSPase or a phytase.

[0043] 31. The method of any one of clauses 1 to 30 wherein the feed composition is administered daily to the animal.

[0044] 32. A method of treating an animal wherein the animal has a disease selected from the group consisting of liver abscess, necrotic laryngitis, calf diphtheria, mastitis, pyometra, and bull nose disease, the method comprising the step of administering to the animal a feed composition or drinking water comprising an effective amount of an additive comprising an isolated Bacillus strain selected from the group consisting of Bacillus strain 86 (NRRL No. B-50944), Bacillus strain 300 (NRRL No. B-50943), Bacillus strain 101 (NRRL No. B-67218), Bacillus strain 235 (NRRL No. B-67219), Bacillus strain 177 (NRRL No. B-67275), Bacillus strain 102 (NRRL No. B-67276), a strain having all of the identifying characteristics of Bacillus strain 86 (NRRL No. B-50944), a strain having all of the identifying characteristics of Bacillus strain 300 (NRRL No. B-50943), a strain having all of the identifying characteristics of Bacillus strain 101 (NRRL No. B-67218), a strain having all of the identifying characteristics of Bacillus strain 235 (NRRL No. B-67219), a strain having all of the identifying characteristics of Bacillus strain 177 (NRRL No. B-67275), a strain having all of the identifying characteristics of Bacillus strain 102 (NRRL No. B-67276), and combinations thereof, wherein the Bacillus strain improves the disease selected from the group consisting of liver abscess, necrotic laryngitis, calf diphtheria, mastitis, pyometra, and bull nose disease.

[0045] 33. The method of clause 32 wherein the animal is selected from the group consisting of a poultry species, a porcine species, a bovine species, an ovine species, a cervid species, an equine species, and a companion animal.

[0046] 34. The method of clause 32 or 33 wherein the animal is a dairy animal.

[0047] 35. The method of clause 34 wherein the dairy animal is selected from the group consisting of a cow, a goat, a sheep, and a buffalo.

[0048] 36. The method of clause 32 or 33 wherein the animal is a deer.

[0049] 37. The method of any one of clauses 32 to 36 wherein the Bacillus strain produces an enzyme selected from the group consisting of an a-galactosidase, a protease, a lipase, an amylase, a xylanase, a cellulase, and combinations thereof.

[0050] 38. The method of any one of clauses 32 to 37 further comprising the step of administering to the animal another bacterial strain selected from the group consisting of another Bacillus strain, a lactic acid bacterial strain, and combinations thereof.

[0051] 39. The method of any one of clauses 32 to 38 wherein Bacillus strain 86(NRRL No. B-50944), Bacillus strain 300 (NRRL No. B-50943), Bacillus strain 101 (NRRLNo. B-67218), Bacillus strain 235 (NRRL No. B-67219), Bacillus strain 177 (NRRL No. B-67275), and Bacillus strain 102 (NRRL No. B-67276) are all administered to the animal.

[0052] 40. The method of any one of clauses 32 to 38 wherein the strain administered is Bacillus strain 101 (NRRL No. B-67218).

[0053] 41. The method of any one of clauses 32 to 38 wherein the strain administered is Bacillus strain 235 (NRRL No. B-67219).

[0054] 42. The method of any one of clauses 32 to 38 wherein the strain administered is Bacillus strain 86 (NRRL No. B-50944).

[0055] 43. The method of any one of clauses 32 to 38 wherein the strain administered is Bacillus strain 300 (NRRL No. B-50943).

[0056] 44. The method of any one of clauses 32 to 38 wherein the strain administered is Bacillus strain 177 (NRRL No. B-67275).

[0057] 45. The method of any one of clauses 32 to 38 wherein the strain administered is Bacillus strain 102 (NRRL No. B-67276).

[0058] 46. The method of any one of clauses 32 to 38 wherein the strains administered are Bacillus strains 102 (NRRL No. B-67276) and 177 (NRRL No. B-67275).

[0059] 47. The method of any one of clauses 32 to 46 wherein the disease is liver abscess.

[0060] 48. The method of any one of clauses 32 to 46 wherein the disease is necrotic laryngitis.

[0061] 49. The method of any one of clauses 32 to 48 wherein the improvement in the disease results in an improvement selected from the group consisting of an improvement in health of the animal, an improvement in carcass quality, an improvement in production efficiency, an improvement in condemnation rate, an improvement in feed efficiency, and combinations thereof.

[0062] 50. The method of any one of clauses 32 to 49 wherein each of the Bacillus strains is administered in the feed composition at a dose of about 1.0 x 103CFU / pound of the feed composition to about 5.0 x 1012CFU / pound of the feed composition.

[0063] 51. The method of any one of clauses 32 to 49 wherein each of the Bacillus strains is administered in the feed composition at a dose of about 0.5 x 107CFU / pound of the feed composition to about 5.0 x 107CFU / pound of the feed composition.

[0064] 52. The method of any one of clauses 32 to 49 wherein each of the Bacillus strains is administered in the feed composition at a dose greater than about 5.0 x 106CFU / pound of the feed composition.

[0065] 53. The method of any one of clauses 32 to 52 further comprising the step of administering an antibiotic to the animal.

[0066] 54. The method of any one of clauses 32 to 53 further comprising the step of administering to the animal an enzyme selected from the group consisting of a galactosidase, a protease, a lipase, an amylase, a hemicellulase, an arabinoxylanase, a xylanase, a cellulase, an NSPase, a phytase, and combinations thereof.

[0067] 55. The method of clause 54 wherein the enzyme is an NSPase or a phytase.

[0068] 56. The method of any one of clauses 32 to 55 wherein the feed composition is administered daily to the animal.

[0069] 57. The method of any one of clauses 1 to 56 wherein the additive is added to the feed composition or drinking water in the form of a concentrate.

[0070] 58. The method of any one of clauses 1 to 56 wherein the additive is added to the feed composition or the drinking water in the form of a superconcentrate.

[0071] 59. The method of any one of clauses 1 to 58 wherein the additive is added to the feed composition or the drinking water in dry form.

[0072] 60. The method of any one of clauses 1 to 59 wherein the feed composition is in pelleted form.

[0073] 61. The method of any one of clauses 1 to 60 wherein the strains are in a form selected from the group consisting of a powder, a liquid, and a pellet form.

[0074] 62. The method of any one of clauses 1 to 61 wherein the additive further comprises a carrier for the Bacillus strains.

[0075] 63. The method of clause 62 wherein the carrier is selected from the group consisting of a bran, rice hulls, a salt, a dextrin, and combinations thereof.

[0076] 64. The method of clause 61 wherein the strains are in powder form.

[0077] 65. The method of clause 61 wherein the strains are in liquid form.

[0078] 66. The method of any one of clauses 1 to 65 wherein the additive further comprises a binder.

[0079] 67. The method of clause 66 wherein the binder is selected from the group consisting of clay, yeast cell wall components, aluminum silicate, glucan, and combinations thereof.BRIEF DESCRIPTION OF THE DRAWINGS

[0080] Fig. 1 shows a photograph of a gel displaying RAPD PCR profiles (Primer 1 to 6) for Bacillus strains 86 and 300. Strain 86 has the top profile and strain 300 has the bottom profile. The leftmost and rightmost lanes have markers and each set of two consecutive lanes between the markers corresponds to Primers 1 to 6 going from left to right.

[0081] Fig. 2 shows a photograph of a gel displaying a RAPD PCR profile (Primer 1 to 6) for Bacillus strain 101. For Primer 1, faint bands appear at approximately 1000, 1500, and greater than 1600 base pairs. For Primer 2, an intense band appears at 650 base pairs and a faint band appears at approximately 400 base pairs. For Primer 3, a faint band appears at approximately 400 base pairs, a faint double band appears at 600 and 700 base pairs, and faint bands appear at 1000 and 1400 base pairs.

[0082] Figs. 3A-3C show photographs of gels displaying a RAPD PCR profile (Primer1 to 3) for Bacillus strain 102. For Primer 1, an intense band appears at 900 base pairs and faint bands appear at 1200 and 1400 base pairs (Fig. 3 A). For Primer 2, intense bands appear at 600, 700, and 1000 base pairs (Fig. 3B). For Primer 3, an intense band appears at 600 base pairs, double band at 700 and 750 base pairs, double band at 900 and 1100 base pairs, and faint bands appear at 400 and greater than 1800 base pairs (Fig. 3C).

[0083] Figs. 4A & 4B show photographs of gels displaying a RAPD PCR profile (Primer 2 and 3) for Bacillus strain 177. For Primer 2, intense bands appear at 600, 700, and 1000 base pairs (Fig. 4A). For Primer 3, intense bands appear at 600 base pairs, double band at 700 and 750 base pairs, double band at 900 and 1100 base pairs, and faint bands appear at 400 and greater than 1800 base pairs (Figs 4B).

[0084] Fig. 5 shows a photograph of a gel displaying a RAPD PCR profile (Primer 1 to 6) for Bacillus strain 235. For Primer 1, faint bands appear at approximately 1000 and greater than 1600 base pairs. For Primer 2, an intense band appears at 650 base pairs and a faint band appears at approximately 400 base pairs. For Primer 3, faint bands appear at approximately 400, 600, 1000, and 1400 base pairs.DETAILED DESCRIPTION OF ILLUSTRATIVE EMBODIMENTS

[0085] Applicants have developed a direct-fed microbial composition that reduces the growth of Fusobacterium necrophorum and Trueperella pyogenes associated with liver abscessand other necrobacillosis, such as necrotic laryngitis (e.g., calf diphtheria), foot rot, and bull nose disease, and other diseases such as lameness, mastitis, metritis, and pyometra,. The direct- fed microbial compositions described herein offer commercial benefits by providing, in a single direct-fed microbial composition, commercially important properties, such as improvement in the disease caused by the Fusobacterium or Trueperella species which results in an improvement selected from the group consisting of an improvement in the health of the animal, an improvement in carcass quality, an improvement in production efficiency, an improvement in condemnation rate, an improvement in feed efficiency, and combinations thereof.

[0086] In various embodiments, the compositions for use in the methods described herein can be a commercial package, a feed additive for an animal feed composition, an additive for the drinking water of an animal, or an animal feed composition (e.g., a complete feed), each comprising one or more of the isolated strains comprising Bacillus strain 86 (NRRL No. B-50944), Bacillus strain 300 (NRRL No. B-50943), Bacillus strain 101 (NRRL No. B- 67218), Bacillus strain 235 (NRRL No. B-67219), Bacillus strain 177 (NRRL No. B-67275), Bacillus strain 102 (NRRL No. B-67276), a strain having all of the identifying characteristics of Bacillus strain 86 (NRRL No. B-50944), a strain having all of the identifying characteristics of Bacillus strain 300 (NRRL No. B-50943), a strain having all of the identifying characteristics of Bacillus strain 101 (NRRL No. B-67218), a strain having all of the identifying characteristics of Bacillus strain 235 (NRRL No. B-67219), a strain having all of the identifying characteristics of Bacillus strain 177 (NRRL No. B-67275), a strain having all of the identifying characteristics of Bacillus strain 102 (NRRL No. B-67276), and combinations thereof.

[0087] In another embodiment, a method is provided of treating an animal wherein the animal has a disease caused by a Fusobacterium or Trueperella species. The method comprises the step of administering to the animal a feed composition or drinking water comprising an effective amount of an additive comprising an isolated Bacillus strain selected from the group consisting of Bacillus strain 86 (NRRL No. B-50944), Bacillus strain 300 (NRRL No. B- 50943), Bacillus strain 101 (NRRL No. B-67218), Bacillus strain 235 (NRRL No. B-67219), Bacillus strain 177 (NRRL No. B-67275), Bacillus strain 102 (NRRL No. B-67276), a strain having all of the identifying characteristics of Bacillus strain 86 (NRRL No. B-50944), a strain having all of the identifying characteristics of Bacillus strain 300 (NRRL No. B-50943), a strain having all of the identifying characteristics of Bacillus strain 101 (NRRL No. B-67218), a strain having all of the identifying characteristics of Bacillus strain 235 (NRRL No. B-67219), a strain having all of the identifying characteristics of Bacillus strain 177 (NRRL No. B-67275), a strain having all of the identifying characteristics of Bacillus strain 102 (NRRL No. B-67276), andcombinations thereof, wherein the Bacillus strain improves the disease caused by the Fusobacterium or Trueperella species.

[0088] In yet another embodiment, a method of treating an animal is provided wherein the animal has a disease selected from the group consisting of liver abscess, necrotic laryngitis, calf diphtheria, mastitis, pyometra, and bull nose disease. The method comprises the step of administering to the animal a feed composition or drinking water comprising an effective amount of an additive comprising an isolated Bacillus strain selected from the group consisting of Bacillus strain 86 (NRRL No. B-50944), Bacillus strain 300 (NRRL No. B-50943), Bacillus strain 101 (NRRL No. B-67218), Bacillus strain 235 (NRRL No. B-67219), Bacillus strain 177 (NRRL No. B-67275), Bacillus strain 102 (NRRL No. B-67276), a strain having all of the identifying characteristics of Bacillus strain 86 (NRRL No. B-50944), a strain having all of the identifying characteristics of Bacillus strain 300 (NRRL No. B-50943), a strain having all of the identifying characteristics of Bacillus strain 101 (NRRL No. B-67218), a strain having all of the identifying characteristics of Bacillus strain 235 (NRRL No. B-67219), a strain having all of the identifying characteristics of Bacillus strain 177 (NRRL No. B-67275), a strain having all of the identifying characteristics of Bacillus strain 102 (NRRL No. B-67276), and combinations thereof, wherein the Bacillus strain improves the disease selected from the group consisting of liver abscess, necrotic laryngitis, calf diphtheria, metritis, pyometra, and bull nose disease.

[0089] The following clauses, and combinations thereof, provide various additional illustrative aspects of the invention described herein. The various embodiments described in any other section of this patent application, including the section titled “BACKGROUND AND SUMMARY OF THE INVENTION” and the EXAMPLES are applicable to any of the following embodiments of the invention described in the numbered clauses below.

[0090] 1. A method of treating an animal wherein the animal has a disease caused by a Fusobacterium or Trueperella species, the method comprising the step of administering to the animal a feed composition or drinking water comprising an effective amount of an additive comprising an isolated Bacillus strain selected from the group consisting of Bacillus strain 86 (NRRL No. B-50944), Bacillus strain 300 (NRRL No. B-50943), Bacillus strain 101 (NRRL No. B-67218), Bacillus strain 235 (NRRL No. B-67219), Bacillus strain 177 (NRRL No. B- 67275), Bacillus strain 102 (NRRL No. B-67276), a strain having all of the identifying characteristics of Bacillus strain 86 (NRRL No. B-50944), a strain having all of the identifying characteristics of Bacillus strain 300 (NRRL No. B-50943), a strain having all of the identifying characteristics of Bacillus strain 101 (NRRL No. B-67218), a strain having all of the identifying characteristics of Bacillus strain 235 (NRRL No. B-67219), a strain having all of the identifying characteristics of Bacillus strain 177 (NRRL No. B-67275), a strain having all of the identifyingcharacteristics of Bacillus strain 102 (NRRL No. B-67276), and combinations thereof, wherein the Bacillus strain improves the disease caused by the Fusobacterium or Trueperella species.

[0091] 2. The method of clause 1 wherein the animal is selected from the group consisting of a poultry species, a porcine species, a bovine species, an ovine species, a cervid species, an equine species, and a companion animal.

[0092] 3. The method of clause 1 or 2 wherein the animal is a dairy animal.

[0093] 4. The method of clause 3 wherein the dairy animal is selected from the group consisting of a cow, a goat, a sheep, and a buffalo.

[0094] 5. The method of clause 1 or 2 wherein the animal is a deer.

[0095] 6. The method of any one of clauses 1 to 5 wherein the Bacillus strain produces an enzyme selected from the group consisting of an a-galactosidase, a protease, a lipase, an amylase, a xylanase, a cellulase, and combinations thereof.

[0096] 7. The method of any one of clauses 1 to 6 further comprising the step of administering to the animal another bacterial strain selected from the group consisting of another Bacillus strain, a lactic acid bacterial strain, and combinations thereof.

[0097] 8. The method of any one of clauses 1 to 7 wherein Bacillus strain 86(NRRL No. B-50944), Bacillus strain 300 (NRRL No. B-50943), Bacillus strain 101 (NRRL No. B-67218), Bacillus strain 235 (NRRL No. B-67219), Bacillus strain 177 (NRRL No. B- 67275), and Bacillus strain 102 (NRRL No. B-67276) are all administered to the animal.

[0098] 9. The method of any one of clauses 1 to 7 wherein the strain administered is Bacillus strain 101 (NRRL No. B-67218).

[0099] 10. The method of any one of clauses 1 to 7 wherein the strain administered is Bacillus strain 235 (NRRL No. B-67219).

[0100] 11. The method of any one of clauses 1 to 7 wherein the strain administered is Bacillus strain 86 (NRRL No. B-50944).

[0101] 12. The method of any one of clauses 1 to 7 wherein the strain administered is Bacillus strain 300 (NRRL No. B-50943).

[0102] 13. The method of any one of clauses 1 to 7 wherein the strain administered is Bacillus strain 177 (NRRL No. B-67275).

[0103] 14. The method of any one of clauses 1 to 7 wherein the strain administered is Bacillus strain 102 (NRRL No. B-67276).

[0104] 15. The method of any one of clauses 1 to 7 wherein the strains administered are Bacillus strains 102 (NRRL No. B-67276) and 177 (NRRL No. B-67275).

[0105] 16. The method of any one of clauses 1 to 15 wherein the disease is caused by a Fusobacterium species.

[0106] 17. The method of clause 16 wherein the Fusobacterium species is selected from the group consisting of Fusobacterium varium, Fusobacterium necrophorum, and combinations thereof.

[0107] 18. The method of clause 16 wherein the Fusobacterium species is aFusoobacterium subspecies selected from the group consisting of Fusobacterium necrophorum necrophorum, Fusobacterium necrophorum funduliforme, and combinations thereof.

[0108] 19. The method of any one of clauses 1 to 15 wherein the disease is caused by a Trueperella species.

[0109] 20. The method of clause 19 wherein the Trueperella species is Trueperella pyogenes.

[0110] 21. The method of any one of clauses 1 to 20 wherein the disease is selected from the group consisting of liver abscess, necrobacillosis, necrotic laryngitis, lameness, mastitis, metritis, pyometra, pneumonia, and combinations thereof.

[0111] 22. The method of clause 21 wherein the necrobacillosis is selected from bull nose disease and foot rot.

[0112] 23. The method of clause 21 wherein the necrotic laryngitis is calf diphtheria.

[0113] 24. The method of any one of clauses 1 to 23 wherein the improvement in the disease caused by the Fusobacterium or Trueperella species results in an improvement selected from the group consisting of an improvement in the health of the animal, an improvement in carcass quality, an improvement in production efficiency, an improvement in condemnation rate, an improvement in feed efficiency, and combinations thereof.

[0114] 25. The method of any one of clauses 1 to 24 wherein each of the Bacillus strains is administered in the feed composition at a dose of about 1.0 x 103CFU / pound of the feed composition to about 5.0 x 1012CFU / pound of the feed composition.

[0115] 26. The method of any one of clauses 1 to 24 wherein each of the Bacillus strains is administered in the feed composition at a dose of about 0.5 x 107CFU / pound of the feed composition to about 5.0 x 107CFU / pound of the feed composition.

[0116] 27. The method of any one of clauses 1 to 24 wherein each of the Bacillus strains is administered in the feed composition at a dose greater than about 5.0 x 106CFU / pound of the feed composition.

[0117] 28. The method of any one of clauses 1 to 27 further comprising the step of administering an antibiotic to the animal.

[0118] 29. The method of any one of clauses 1 to 28 further comprising the step of administering to the animal an enzyme selected from the group consisting of a galactosidase, aprotease, a lipase, an amylase, a hemicellulase, an arabinoxylanase, a xylanase, a cellulase, an NSPase, a phytase, and combinations thereof.

[0119] 30. The method of clause 29 wherein the enzyme is an NSPase or a phytase.

[0120] 31. The method of any one of clauses 1 to 30 wherein the feed composition is administered daily to the animal.

[0121] 32. A method of treating an animal wherein the animal has a disease selected from the group consisting of liver abscess, necrotic laryngitis, calf diphtheria, mastitis, pyometra, and bull nose disease, the method comprising the step of administering to the animal a feed composition or drinking water comprising an effective amount of an additive comprising an isolated Bacillus strain selected from the group consisting of Bacillus strain 86 (NRRL No. B-50944), Bacillus strain 300 (NRRL No. B-50943), Bacillus strain 101 (NRRL No. B-67218), Bacillus strain 235 (NRRL No. B-67219), Bacillus strain 177 (NRRL No. B-67275), Bacillus strain 102 (NRRL No. B-67276), a strain having all of the identifying characteristics of Bacillus strain 86 (NRRL No. B-50944), a strain having all of the identifying characteristics of Bacillus strain 300 (NRRL No. B-50943), a strain having all of the identifying characteristics of Bacillus strain 101 (NRRL No. B-67218), a strain having all of the identifying characteristics of Bacillus strain 235 (NRRL No. B-67219), a strain having all of the identifying characteristics of Bacillus strain 177 (NRRL No. B-67275), a strain having all of the identifying characteristics of Bacillus strain 102 (NRRL No. B-67276), and combinations thereof, wherein the Bacillus strain improves the disease selected from the group consisting of liver abscess, necrotic laryngitis, calf diphtheria, mastitis, pyometra, and bull nose disease.

[0122] 33. The method of clause 32 wherein the animal is selected from the group consisting of a poultry species, a porcine species, a bovine species, an ovine species, a cervid species, an equine species, and a companion animal.

[0123] 34. The method of clause 32 or 33 wherein the animal is a dairy animal.

[0124] 35. The method of clause 34 wherein the dairy animal is selected from the group consisting of a cow, a goat, a sheep, and a buffalo.

[0125] 36. The method of clause 32 or 33 wherein the animal is a deer.

[0126] 37. The method of any one of clauses 32 to 36 wherein the Bacillus strain produces an enzyme selected from the group consisting of an a-galactosidase, a protease, a lipase, an amylase, a xylanase, a cellulase, and combinations thereof.

[0127] 38. The method of any one of clauses 32 to 37 further comprising the step of administering to the animal another bacterial strain selected from the group consisting of another Bacillus strain, a lactic acid bacterial strain, and combinations thereof.

[0128] 39. The method of any one of clauses 32 to 38 wherein Bacillus strain 86(NRRL No. B-50944), Bacillus strain 300 (NRRL No. B-50943), Bacillus strain 101 (NRRLNo. B-67218), Bacillus strain 235 (NRRL No. B-67219), Bacillus strain 177 (NRRL No. B- 67275), and Bacillus strain 102 (NRRL No. B-67276) are all administered to the animal.

[0129] 40. The method of any one of clauses 32 to 38 wherein the strain administered is Bacillus strain 101 (NRRL No. B-67218).

[0130] 41. The method of any one of clauses 32 to 38 wherein the strain administered is Bacillus strain 235 (NRRL No. B-67219).

[0131] 42. The method of any one of clauses 32 to 38 wherein the strain administered is Bacillus strain 86 (NRRL No. B-50944).

[0132] 43. The method of any one of clauses 32 to 38 wherein the strain administered is Bacillus strain 300 (NRRL No. B-50943).

[0133] 44. The method of any one of clauses 32 to 38 wherein the strain administered is Bacillus strain 177 (NRRL No. B-67275).

[0134] 45. The method of any one of clauses 32 to 38 wherein the strain administered is Bacillus strain 102 (NRRL No. B-67276).

[0135] 46. The method of any one of clauses 32 to 38 wherein the strains administered are Bacillus strains 102 (NRRL No. B-67276) and 177 (NRRL No. B-67275).

[0136] 47. The method of any one of clauses 32 to 46 wherein the disease is liver abscess.

[0137] 48. The method of any one of clauses 32 to 46 wherein the disease is necrotic laryngitis.

[0138] 49. The method of any one of clauses 32 to 48 wherein the improvement in the disease results in an improvement selected from the group consisting of an improvement in health of the animal, an improvement in carcass quality, an improvement in production efficiency, an improvement in condemnation rate, an improvement in feed efficiency, and combinations thereof.

[0139] 50. The method of any one of clauses 32 to 49 wherein each of the Bacillus strains is administered in the feed composition at a dose of about 1.0 x 103CFU / pound of the feed composition to about 5.0 x 1012CFU / pound of the feed composition.

[0140] 51. The method of any one of clauses 32 to 49 wherein each of the Bacillus strains is administered in the feed composition at a dose of about 0.5 x 107CFU / pound of the feed composition to about 5.0 x 107CFU / pound of the feed composition.

[0141] 52. The method of any one of clauses 32 to 49 wherein each of the Bacillus strains is administered in the feed composition at a dose greater than about 5.0 x 106CFU / pound of the feed composition.

[0142] 53. The method of any one of clauses 32 to 52 further comprising the step of administering an antibiotic to the animal.

[0143] 54. The method of any one of clauses 32 to 53 further comprising the step of administering to the animal an enzyme selected from the group consisting of a galactosidase, a protease, a lipase, an amylase, a hemicellulase, an arabinoxylanase, a xylanase, a cellulase, an NSPase, a phytase, and combinations thereof.

[0144] 55. The method of clause 54 wherein the enzyme is an NSPase or a phytase.

[0145] 56. The method of any one of clauses 32 to 55 wherein the feed composition is administered daily to the animal.

[0146] 57. The method of any one of clauses 1 to 56 wherein the additive is added to the feed composition or drinking water in the form of a concentrate.

[0147] 58. The method of any one of clauses 1 to 56 wherein the additive is added to the feed composition or the drinking water in the form of a superconcentrate.

[0148] 59. The method of any one of clauses 1 to 58 wherein the additive is added to the feed composition or the drinking water in dry form.

[0149] 60. The method of any one of clauses 1 to 59 wherein the feed composition is in pelleted form.

[0150] 61. The method of any one of clauses 1 to 60 wherein the strains are in a form selected from the group consisting of a powder, a liquid, and a pellet form.

[0151] 62. The method of any one of clauses 1 to 61 wherein the additive further comprises a carrier for the Bacillus strains.

[0152] 63. The method of clause 62 wherein the carrier is selected from the group consisting of a bran, rice hulls, a salt, a dextrin, and combinations thereof.

[0153] 64. The method of clause 61 wherein the strains are in powder form.

[0154] 65. The method of clause 61 wherein the strains are in liquid form.

[0155] 66. The method of any one of clauses 1 to 65 wherein the additive further comprises a binder.

[0156] 67. The method of clause 66 wherein the binder is selected from the group consisting of clay, yeast cell wall components, aluminum silicate, glucan, and combinations thereof.

[0157] In various embodiments, the animal to which a feed additive, a feed composition, or drinking water as described herein is administered can be selected from the group consisting of a poultry species, a porcine species, a bovine species, an ovine species, an equine species, a cervid species, and a companion animal. In the embodiment where the animal is a companion animal, the companion animal can be, for example, a canine species or a feline species. In the embodiment where the animal is a porcine species, the porcine species can be selected from the group consisting of a grow finish pig, a nursery pig, a sow, and a breeding stock pig. In various exemplary embodiments, the animal can be selected from the group consisting of a chicken (e.g., a broiler or a layer), a pig, a horse, a pony, a cow, a buffalo, a deer, a turkey, a goat, a sheep, a quail, a pheasant, an ostrich, a duck, and combinations thereof. In one illustrative embodiment, the animal can be a cow. In another illustrative embodiment, the animal can be a deer. In another illustrative embodiment, the animal can be a goat. In another illustrative embodiment, the animal can be a sheep. In another illustrative embodiment, the animal can be a pig. In yet another embodiment, the animal can be a dairy animal, or another ruminant.

[0158] In any embodiments described herein, except those requiring administering all of the strains in combination, the strains can be administered alone or in any combination. In one embodiment, an effective amount of the Bacillus strain can be administered to the animal. By “effective amount” is meant an amount of the Bacillus strain (e.g., strain 86, 300, 101, or 235, 177, or 102, or strains having all of their identifying characteristics) capable of improving a disease (e.g., liver abscess, necrobacillosis, necrotic laryngitis, foot rot, bull nose disease, calf diphtheria, lameness, mastitis, metritis, pyometra, pneumonia, and combinations thereof), improving a disease caused by a Fusobacterium or Trueperella species, or capable of causing an improvement selected from the group consisting of an improvement in the health of the animal, an improvement in carcass quality, an improvement in production efficiency, an improvement in condemnation rate, an improvement in feed efficiency, and combinations thereof, by any mechanism.

[0159] In another illustrative aspect, the disease that is improved can be caused by a Fusobacterium or Trueperella species, wherein the Fusobacterium species is selected from the group consisting of Fusobacterium varium, Fusobacterium necrophorum, and combinations thereof. In another aspect, the disease that is improved can be caused by a Fusobacterium species that is a Fusoobacterium subspecies selected from the group consisting of Fusobacterium necrophorum necrophorum, Fusobacterium necrophorum funduliforme, and combinations thereof. In yet another embodiment, the disease that is improved can be caused by a Trueperella species, and the Trueperella species can be Trueperella pyogenes.

[0160] As used herein, “improve a disease or disorder” can mean reducing the number of animals with the disease or disorder, preventing the disease or disorder, reducing the symptoms of the disease or disorder in animals, inhibiting the growth of a pathogen such as a Fusobacterium or Trueperella species, or combinations thereof, wherein the disease or disorder is selected from the group consisting of liver abscess, necrobacillosis, necrotic laryngitis, foot rot, bull nose disease, calf diphtheria, lameness, mastitis, metritis, pyometra, pneumonia, and combinations thereof. As used herein, the terms “inhibit”, “inhibiting”, “inhibition”, “decrease”, “decreasing”, “increase”, “increasing”, “improve”, “improving”, “improvement”, “reduce”, or “reducing” for example, are all relative to the condition of the animals before administering the strain or strains to the animals or, for example, relative to an animal not fed the Bacillus strain. As used herein, “treating”, “treat”, and “treatment” for animals can mean, for example, treating a disease in the animal with the Bacillus strains described herein, feeding the animal the Bacillus strains described herein, or administering the Bacillus strains described herein to the animal.

[0161] In embodiments described herein wherein the compositions of the present invention comprising Bacillus strains 86, and / or 300, and / or 101, and / or 235, and / or 177, and / or 102, and / or strains having all of their identifying characteristics, are administered to an animal, the compositions are preferably administered to animals orally in a feed composition or in drinking water, but any other effective method of administration known to those skilled in the art may be utilized. In one illustrative embodiment, the Bacillus strains 86, and / or 300, and / or 101, and / or 235, and / or 177, and / or 102, and / or strains having all of their identifying characteristics, are provided in the form of an additive for addition to the drinking water of the animal.

[0162] In another illustrative embodiment, the Bacillus strains 86, and / or 300, and / or 101, and / or 235, and / or 177, and / or 102, and / or strains having all of their identifying characteristics, are provided in the form of a feed additive for addition to a feed composition. The feed composition may contain Bacillus strain 86, and / or 300, and / or 101, and / or 235, and / or 177, and / or 102, and / or strains having all of their identifying characteristics, in a mixture with an animal feed blend, including any art-recognized animal feed blend or any animal feed blend described herein. As used herein, “feed composition” or “animal feed composition” means a feed composition comprising Bacillus strain 86, and / or 300, and / or 101, and / or 235, and / or 177, and / or 102, and / or strains having all of their identifying characteristics, in a mixture with an animal feed blend, and, optionally any other components that could be used in a feed composition, including other bacterial strains, such as other Bacillus strains or Lactobacillus strains. The feed additive, feed composition, animal feed composition, or additive for drinkingwater can comprise all, or one or more, of strains 86, and / or 300, and / or 101, and / or 235, and / or 177, and / or 102, and / or strains having all of their identifying characteristics. In one embodiment, the feed additive, feed composition, animal feed composition, or additive for drinking water comprises Bacillus strains 102 (NRRL No. B-67276) and 177 (NRRL No. B- 67275).

[0163] Any animal feed blend, including those known in the art and those described herein, may be used in accordance with the methods and compositions described in this patent application, such as rapeseed meal, cottonseed meal, soybean meal, cornmeal, barley, wheat, silage, and haylage. In various embodiments, a corn silage and alfalfa hay-based total mixed ration or a corn silage mixed ration can be used. In various embodiments, the animal feed blend can be supplemented with Bacillus strain 86, and / or Bacillus strain 300, and / or Bacillus strain 101, and / or Bacillus strain 235, and / or Bacillus strain 177, and / or Bacillus strain 102, and / or strains having all of their identifying characteristics, but other ingredients may optionally be added to the animal feed blend, including other different bacterial strains, such as other Bacillus strains or Lactobacillus strains.

[0164] In various illustrative embodiments, optional ingredients of the animal feed blend include sugars and complex carbohydrates such as both water-soluble and water-insoluble monosaccharides, disaccharides, and polysaccharides. Other optional ingredients include dried distillers grain solubles, fat (e.g., crude fat), phosphorous, sodium bicarbonate, limestone, salt, phytate, calcium, sodium, sulfur, magnesium, potassium, copper, iron, manganese, zinc, ash, fish oil, an oil derived from fish meal, raw seed (e.g., flaxseed), an antioxidant, and starch. In another embodiment, minerals may be added in the form of a mineral premix.

[0165] Optional amino acid ingredients that may be added to the animal feed blend are arginine, histidine, isoleucine, leucine, lysine, cysteine, methionine, phenylalanine, threonine, tryptophan, valine, tyrosine ethyl HC1, alanine, aspartic acid, sodium glutamate, glycine, proline, serine, cysteine ethyl HC1, and analogs, and salts thereof. Vitamins that may be optionally added are thiamine HC1, riboflavin, pyridoxine HC1, niacin, niacinamide, inositol, choline chloride, calcium pantothenate, biotin, folic acid, ascorbic acid, and vitamins A, B, K, D, E, and the like. In another embodiment, vitamins may be added in the form of a vitamin premix. In yet another embodiment, protein ingredients may be added to the animal feed blend and include protein obtained from meat meal, bone meal, or fish meal, liquid or powdered egg, fish solubles, crude protein, and the like.

[0166] In another illustrative aspect, any medicament ingredients known in the art may be added to the animal feed blend or to an additive for the drinking water of the animal, such asantibiotics. In another embodiment, the animal feed blend, the feed composition, the feed additive, or the additive for the drinking water of the animal may or may not contain antibiotics.

[0167] In another illustrative embodiment, one or more enzymes may be added to the animal feed blend. In various embodiments, the enzymes that may be added include a galactosidase, a phytase, a protease, a lipase, an amylase, a hemicellulase, an arabinoxylanase, a xylanase, a cellulase, an NSPase, combinations thereof, and any other enzyme that improves the effectiveness of the feed composition for the uses described herein. In yet another embodiment, yeast, fungi (e.g., Aspergillus or Trichodermd), or micronutrients may be added to the animal feed. Any of the ingredients described above that are suitable for addition to an additive for the drinking water of the animal may be added as a component of the additive for the drinking water of the animal as described herein.

[0168] In various illustrative embodiments, the Bacillus strain (e.g., Bacillus strain 86, and / or 300, and / or 101, and / or 235, and / or 177, and / or 102, and / or strains having all of their identifying characteristics), or any other different bacterial strains added in addition to these Bacillus strains, can be administered in the feed composition, each strain at a dose of about 1.0 x 103CFU / pound of the feed composition to about 5.0 x 1012CFU / pound of the feed composition, each strain at a dose of about 0.5 x 107CFU / pound of the feed composition to about 5.0 x 107CFU / pound of the feed composition, or at a dose greater than about 5.0 x 106CFU / pound of the feed composition. In other embodiments, each Bacillus strain (e.g., Bacillus strain 86, and / or 300, and / or 101, and / or 235, and / or 177, and / or 102, and / or strains having all of their identifying characteristics) can be administered in the feed composition at a dose greater than about 1.0 x 103CFU / pound of the feed composition, at a dose greater than about 1.1 x 103CFU / pound of the feed composition, at a dose greater than about 1.25 x 103CFU / pound of the feed composition, at a dose greater than about 1.5 x 103CFU / pound of the feed composition, at a dose greater than about 1.75 x 103CFU / pound of the feed composition, at a dose greater than about 1.0 x 104CFU / pound of the feed composition, at a dose greater than about 2.0 x 104CFU / pound of the feed composition, at a dose greater than about 3.0 x 104CFU / pound of the feed composition, at a dose greater than about 4.0 x 104CFU / pound of the feed composition, at a dose greater than about 5.0 x 104CFU / pound of the feed composition, at a dose greater than about 6.0 x 104CFU / pound of the feed composition, at a dose greater than about 7.0 x 104CFU / pound of the feed composition, at a dose greater than about 8.0 x 104CFU / pound of the feed composition, at a dose greater than about 1.0 x 105CFU / pound of the feed composition, at a dose greater than about 1.0 x 106CFU / pound of the feed composition, at a dose greater than about 2.0 x 106CFU / pound of the feed composition, at a dose greater than about 3.0 x 106CFU / pound of the feed composition, at a dose greater than about 4.0 x 106CFU / pound of the feed composition, at a dose greater than about 5.0 x 106CFU / pound of the feed composition, at a dose greater than about 6.0 x 106CFU / pound of the feed composition, at a dose greater than about 7.0 x 106CFU / pound of the feed composition, at a dose greater than about 8.0 x 106CFU / pound of the feed composition, at a dose greater than about 9.0 x 106CFU / pound of the feed composition, at a dose greater than about 1 .0 x 107CFU / pound of the feed composition, at a dose greater than about 2.0 x 107CFU / pound of the feed composition, at a dose greater than about 3.0 x 107CFU / pound of the feed composition, at a dose greater than about 4.0 x 107CFU / pound of the feed composition, at a dose greater than about 5.0 x 107CFU / pound of the feed composition, at a dose greater than about 1.0 x 108CFU / pound of the feed composition, at a dose greater than about 1.0 x 109CFU / pound of the feed composition, at a dose greater than about 1.0 x 1010CFU / pound of the feed composition, at a dose greater than about 1.0 x 1011CFU / pound of the feed composition, or at a dose greater than about 1.0 x 1012CFU / pound of the feed composition. In all of the embodiments described in this paragraph, the animal can be fed, or can consume, about 30 pounds, about 35 pounds, about 40 pounds, about 45 pounds, about 50 pounds, about 55 pounds, about 60 pounds, about 65 pounds, or about 70 pounds of the feed composition per day.

[0169] In other embodiments, the Bacillus strains (e.g., Bacillus strain 86, 300, 101, 235, 177, and 102, and / or strains having all of their identifying characteristics), can he administered in the feed composition by adding about 15 grams of a feed additive to the complete feed composition wherein the 15 grams of the additive comprises all of the strains to provide a total of the combined strains in the 15 grams of the additive of about 1.0 x 109CFU, about 2.0 x 109CFU, about 3.0 x 109CFU, about 4.0 x 109CFU, about 5.0 x 109CFU, about 6.0 x 109CFU, about 7.0 x 109CFU, about 8.0 x 109CFU, about 9.0 x 109CFU, or about 1.0 x 1010CFU to be fed to the animal per day. In all of the embodiments described in this paragraph, the animal can be fed, or can consume, about 30 pounds, about 35 pounds, about 40 pounds, about 45 pounds, about 50 pounds, about 55 pounds, about 60 pounds, about 65 pounds, or about 70 pounds of the complete feed composition per day, and the 15 grams of the feed additive can be added to any of these amounts of the feed composition.

[0170] In another embodiment, the Bacillus strains (e.g., Bacillus strain 86, 300, 101, 235, 177, and 102, and / or strains having all of their identifying characteristics), can be administered in the feed composition by adding about 15 grams of a feed additive comprising about 7.0 x 109CFU total of the combined strains (e.g., about 7.35 x 109CFU total) to, e.g., about 30 pounds, about 35 pounds, about 40 pounds, about 45 pounds, about 50 pounds, about 55 pounds, about 60 pounds, about 65 pounds, or about 70 pounds of the complete feed composition to be fed, or consumed, per animal per day.

[0171] In any of the dose embodiments for Bacillus strains 86, 300, 101, 235, 177, and 102, and / or strains having all of their identifying characteristics, described above, the Bacillus strains (Bacillus strain 86, 300, 101, 235, 177, and 102, and / or strains having all of their identifying characteristics) can be included in the feed additive, for addition to the feed composition, in exemplary ratios selected from any of the ratios shown below, wherein the ratios refer to the Bacillus strains 86, 300, 101, 235, 177, and 102, and / or strains having all of their identifying characteristics, in that order.1:1:1:1:1:1 l:0.8:l:l:l:l1: 1:0.8: 1: 1: 11: 1: 1:0.8: 1: 1 l:l:l:l:0.8:l l:l:l:l:l:0.8 l:0.5:l:l:l:l1: 1:0.5: 1: 1: 11:1: 1:0.5: 1: 11 :1 :1 : 1 :0.5:l l:l:l: l:l:0.5 l:0.2:l:l: l:l l:l:0.2:l: l:l l:l:l:0.2: l:l l:l:l: l:0.2:l l:l:l: l:l:0.20.8:0.5:l:l:l: 1 l:0.8:0.5:l:l: 1 l:l:0.8:0.5:l: 1 l:l:l:0.8:0.5: l1:1:1: l:0.8:0.50.2:0.5: l: l: l: 1 l:0.2:0.5:l:l: 1 l:l:0.2:0.5:l: l l:l:l:0.2:0.5: l l:l:l: l:0.2:0.50.8:0.2:l:l:l: 1l:0.8:0.2:l:l: l l:l:0.8:0.2:l:l l:l:l:0.8:0.2:l 1: 1 : 1: l:0.8:0.2 0.8:0.5:0.2: 1 :1 :1 l:0.8:0.5:0.2:l:l l:l:0.8:0.5:0.2:l l:l:l:0.8:0.5:0.2 0.2:l:l:l:0.8:0.5 0.8:0.2:0.5: l:l:l l:0.8:0.2:0.5:l:l l:l:0.8:0.2:0.5:l l:l:l:0.8:0.2:0.5 0.5:l:l:l:0.8:0.2 0.2:0.8.:0.5:l:l:l l:0.2:0.8:0.5:l:l l:l:0.2:0.8:0.5:l 1 :1 :l :0.2:0.8:0.5 0.2:l:l:l:0.8:0.5

[0172] In various embodiments, the Bacillus strain (e.g., Bacillus strain 86, and / or 300, and / or 101, and / or 235, and / or 177, and / or 102) for use in accordance with the methods and compositions described herein can be selected from the group consisting of Bacillus strain 86 (NRRL No. B-50944), Bacillus strain 300 (NRRL No. B-50943), Bacillus strain 101 (NRRL No. B-67218), Bacillus strain 235 (NRRL No. B-67219), Bacillus strain 177 (NRRL No. B- 67275), Bacillus strain 102 (NRRL No. B-67276), a strain having all of the identifying characteristics of Bacillus strain 86 (NRRL No. B-50944), a strain having all of the identifying characteristics of Bacillus strain 300 (NRRL No. B-50943), a strain having all of the identifying characteristics of Bacillus strain 101 (NRRL No. B-67218), a strain having all of the identifying characteristics of Bacillus strain 235 (NRRL No. B-67219), a strain having all of the identifying characteristics of Bacillus strain 177 (NRRL No. B-67275), and a strain having all of the identifying characteristics of Bacillus strain 102 (NRRL No. B-67276). Bacillus strain MDG 101 and Bacillus strain MDG 235 were deposited on January 4, 2016 at the Agricultural Research Service Culture Collection (NRRL), National Center for Agricultural UtilizationResearch, Agricultural Research Service, USDA, 1815 North University Street, Peoria, Illinois61604-3999, and were given accession numbers B-67218 and B-67219, respectively. Bacillus strain MGL177 and Bacillus strain MGL102 were deposited on June 7, 2016 at the Agricultural Research Service Culture Collection (NRRL), National Center for Agricultural Utilization Research, Agricultural Research Service, USDA, 1815 North University Street, Peoria, Illinois 61604-3999, and were given accession numbers B-67275 and B-67276, respectively. Bacillus strain MDG86 and Bacillus strain MDG300 were deposited on March 14, 2014 at the Agricultural Research Service Culture Collection (NRRL), National Center for Agricultural Utilization Research, Agricultural Research Service, USDA, 1815 North University Street, Peoria, Illinois 61604-3999, and were given accession numbers B-50944 and B-50943, respectively. The NRRL strain designations MDG86 and MDG300 are equivalent to Bacillus strain 86 and 300, respectively, as referred to in the application. The NRRL strain designations MDG 101, MDG 235, MGL177, and MGL102 are equivalent to Bacillus strain 101, 235, 177, and 102 respectively, as referred to in the application. The deposits were made under the provisions of the Budapest Treaty on the International Recognition of the Deposit of Microorganisms for the Purposes of Patent Procedure. The strains described herein can be referred to as Bacillus strain 86, Bacillus strain 300, Bacillus strain 101, Bacillus strain 235, Bacillus strain 177, or Bacillus strain 102 or strain 86, strain 300, strain 101, strain 235, strain 177, or strain 102.

[0173] Bacillus strains 102 and 177 are Bacillus pumilus strains, Bacillus strain 77 is a Bacillus licheniformis strain, and all other strains described in the application are Bacillus subtilus strains.

[0174] Any of these strains can be administered alone or in combination in the form of a feed composition (e.g., a complete feed comprising an animal feed blend) or drinking water for an animal. In one embodiment, multiple strains are administered in combination in a single composition. In another embodiment, multiple strains are administered in separate compositions.

[0175] In another embodiment, one or more of the Bacillus strains described in the preceding paragraphs (e.g., Bacillus strain 86, and / or Bacillus strain 300, and / or Bacillus strain 101, and / or Bacillus strain 235, and / or Bacillus strain 177, and / or Bacillus strain 102, and / or strains having all of their identifying characteristics) can be administered to the animal along with another different bacterial strain selected from the group consisting of another Bacillus strain, a lactic acid bacterial strain, and combinations thereof. In yet another embodiment, one or more of the Bacillus strains described in the preceding paragraphs (e.g., Bacillus strain 86, and / or Bacillus strain 300, and / or Bacillus strain 101, and / or Bacillus strain 235, and / or Bacillus strain 177, and / or Bacillus strain 102, and / or strains having all of their identifyingcharacteristics) can be administered to the animal along with any other different bacterial strain effective for the uses described herein.

[0176] As used herein “a strain having all of the identifying characteristics of’ Bacillus strain 86, Bacillus strain 300, Bacillus strain 101, Bacillus strain 235, Bacillus strain 177, or Bacillus strain 102 can be a mutant strain having all of the identifying characteristics of Bacillus strain 86, Bacillus strain 300, Bacillus strain 101, Bacillus strain 235, Bacillus strain 177, or Bacillus strain 102; e.g., having a DNA fingerprint based on DNA analysis that corresponds to the DNA fingerprint of Bacillus strain 86, Bacillus strain 300, Bacillus strain 101, Bacillus strain 235, Bacillus strain 177, or Bacillus strain 102, enzyme activities that correspond to Bacillus strain 86, Bacillus strain 300, Bacillus strain 101, Bacillus strain 235, Bacillus strain 177, or Bacillus strain 102, antimicrobial activity that corresponds to Bacillus strain 86, Bacillus strain 300, Bacillus strain 101, Bacillus strain 235, Bacillus strain or 177, or Bacillus strain 102, antibiotic sensitivity and tolerance profiles that correspond to Bacillus strain 86, Bacillus strain 300, Bacillus strain 101, Bacillus strain 235, Bacillus strain 177, or Bacillus strain 102). In alternate embodiments, the mutation can be a natural mutation, or a genetically engineered mutation. In another embodiment, “a strain having all of the identifying characteristics of” Bacillus strain 86, Bacillus strain 300, Bacillus strain 101, Bacillus strain 235, Bacillus strain 177, or Bacillus strain 102 can be a strain, for example, produced by isolating one or more plasmids from Bacillus strain 86, Bacillus strain 300, Bacillus strain 101, Bacillus strain 235, Bacillus strain 177, or Bacillus strain 102 and introducing the one or more plasmids into another bacterium, such as another Bacillus strain, as long as the one or more plasmids contain DNA that provides the identifying characteristics of Bacillus strain 86, Bacillus strain 300, Bacillus strain 101, Bacillus strain 235, Bacillus strain 177, or Bacillus strain 102 (e.g., a DNA fingerprint based on DNA analysis that corresponds to the DNA fingerprint of Bacillus strain 86, Bacillus strain 300, Bacillus strain 101, Bacillus strain 235, Bacillus strain 177, or Bacillus strain 102).

[0177] The feed composition or drinking water comprising Bacillus strain 86, and / or Bacillus strain 300, and / or Bacillus strain 101, and / or Bacillus strain 235, and / or Bacillus strain 177, and / or Bacillus strain 102, and / or strains having all of their identifying characteristics, may be administered to the animal for any time period that is effective for the uses described herein, or combinations thereof. For example, in one embodiment the feed composition or drinking water may be provided to the animal daily. In an alternate embodiment, the feed composition or drinking water may be administered to the animal during lactation and / or during gestation. The time periods for administration of the feed composition or drinking water described above are non- limiting examples and it should be appreciated that any time period or administrationschedule determined to be effective for the uses described herein, or combinations thereof, may be used.

[0178] In additional embodiments of the invention, compositions comprising Bacillus strain 86, Bacillus strain 300, Bacillus strain 101, Bacillus strain 235, Bacillus strain 177, and Bacillus strain 102, and / or strains having all of their identifying characteristics, are provided. In one embodiment, a commercial package for animal use is provided comprising one or more or all of the isolated strains comprising Bacillus strain 86 (NRRL No. B-50944), Bacillus strain 300 (NRRL No. B-50943), Bacillus strain 101 (NRRL No. B-67218), Bacillus strain 235 (NRRL No. B-67219), Bacillus strain 177 (NRRL No. B-67275), and Bacillus strain 102 (NRRL No. B-67276).

[0179] In another embodiment, a feed additive for an animal feed is provided comprising one or more or all of the isolated strains comprising Bacillus strain 86 (NRRL No. B-50944), Bacillus strain 300 (NRRL No. B-50943), Bacillus strain 101 (NRRL No. B-67218), Bacillus strain 235 (NRRL No. B-67219), Bacillus strain 177 (NRRL No. B-67275), and Bacillus strain 102 (NRRL No. B-67276).

[0180] In yet another embodiment, an additive for the drinking water of an animal is provided comprising one or more or all of the isolated strains comprising Bacillus strain 86 (NRRL No. B-50944), Bacillus strain 300 (NRRL No. B-50943), Bacillus strain 101 (NRRL No. B-67218), Bacillus strain 235 (NRRL No. B-67219), Bacillus strain 177 (NRRL No. B- 67275), and Bacillus strain 102 (NRRL No. B-67276).

[0181] In still another embodiment, an animal feed composition is provided comprising one or more or all of the isolated strains comprising Bacillus strain 86 (NRRL No. B-50944), Bacillus strain 300 (NRRL No. B-50943), Bacillus strain 101 (NRRL No. B-67218), Bacillus strain 235 (NRRL No. B-67219), Bacillus strain 177 (NRRL No. B-67275), and Bacillus strain 102 (NRRL No. B-67276).

[0182] In one embodiment, the feed additive for addition to an animal feed blend to produce a complete feed composition can be mixed with the animal feed blend, for example, with an automated micro-nutrient delivery system, or, for example, by hand- weighing and addition to achieve any of the doses of Bacillus strain 86, Bacillus strain 300, Bacillus strain 101, Bacillus strain 235, Bacillus strain 177, or Bacillus strain 102, and / or strains having all of their identifying characteristics, described herein, for administration to the animal in the form of a complete feed composition. The mixing can also be done by any other suitable method known in the art for combining direct-fed microbials with an animal feed blend to obtain a uniform mixture. In various embodiments, the mixing can be done for any suitable time period (e.g., about 1 to about 4 minutes). In the embodiment where Bacillus strain 86, Bacillus strain300, Bacillus strain 101, Bacillus strain 235, Bacillus strain 177, and Bacillus strain 102, and / or strains having all of their identifying characteristics, are in the form of an additive for the drinking water of the animal, Bacillus strain 86, Bacillus strain 300, Bacillus strain 101, Bacillus strain 235, Bacillus strain 177, and Bacillus strain 102, and / or strains having all of their identifying characteristics, can he in the form of, for example, a powder, a liquid, or pellets, and can be mixed with the drinking water using any suitable method known in the art to achieve any of the doses of Bacillus strain 86, Bacillus strain 300, Bacillus strain 101, Bacillus strain 235, Bacillus strain 177, or Bacillus strain 102, and / or strains having all of their identifying characteristics, described herein, for administration to the animal in the drinking water of the animal. Bacillus strain 86, Bacillus strain 300, Bacillus strain 101, Bacillus strain 235, Bacillus strain 177, and Bacillus strain 102, and / or strains having all of their identifying characteristics, can also be fed directly to the animal orally (i.e., by oral insertion) in the form of a powder, a liquid, a capsule, a top-dressing, a gel, or a pellet.

[0183] In any of the composition embodiments described herein, Bacillus strain 86, Bacillus strain 300, Bacillus strain 101, Bacillus strain 235, Bacillus strain 177, and Bacillus strain 102, and / or strains having all of their identifying characteristics, can cause an effect selected from the group consisting of improving a disease (e.g., liver abscess, necrobacillosis, necrotic laryngitis, foot rot, bull nose disease, calf diphtheria, lameness, mastitis, metritis, pyometra, pneumonia, and combinations thereof), improving a disease caused by a Fusobacterium or Trueperella species, or causing an improvement selected from the group consisting of an improvement in the health of the animal, an improvement in carcass quality, an improvement in production efficiency, an improvement in condemnation rate, an improvement in feed efficiency, and combinations thereof, by any mechanism.

[0184] In one illustrative aspect, the feed additive, additive for the drinking water of the animal, or the feed composition can be in the form of a commercial package. In another illustrative embodiment, the feed additive or additive for the drinking water of the animal comprising Bacillus strain 86, Bacillus strain 300, Bacillus strain 101, Bacillus strain 235, Bacillus strain 177, and Bacillus strain 102, and / or strains having all of their identifying characteristics, in the commercial package can be in the form of a concentrate comprising, e.g., about 1 x 10sto about 7 x IO10CFU / g of the strains in combination, or a superconcentrate (e.g., about 8 x 1010to about 5 x 1012CFU / g of the strains in combination). In another embodiment, the feed additive, feed composition, or additive for the drinking water of the animal, or the Bacillus strain 86, Bacillus strain 300, Bacillus strain 101, Bacillus strain 235, Bacillus strain 177, and Bacillus strain 102, and / or strains having all of their identifying characteristics, are in a composition in a commercial package, and can be in a dry form (e.g., a powder), a pelletedform, a liquid form, in the form of a top-dressing, or in the form of a gel, in the form of a capsule, or any other suitable form.

[0185] In yet another embodiment, the strains in the form of a commercial package can be, for example, in a dry form (e.g., a powder or freeze-dried form), in a pelleted form, or in a liquid form.

[0186] In another illustrative embodiment, the commercial package, feed additive, additive for the drinking water of the animal, or feed composition can further comprise a carrier for the Bacillus strain 86, Bacillus strain 300, Bacillus strain 101, Bacillus strain 235, Bacillus strain 177, and Bacillus strain 102, and / or strains having all of their identifying characteristics,. The carrier can be selected from the group consisting of a bran, rice hulls, a salt, mineral oil, a dextrin (e.g., maltodextrin), whey, sugar, limestone, dried starch, sodium silico aluminate, vegetable oil, and combinations thereof. In another embodiment, the carrier can be any suitable carrier known in the art for a direct- fed microbial. In another embodiment, the commercial package, feed additive, additive for the drinking water of the animal, or feed composition can further comprise a binder such as clay, yeast cell wall components, aluminum silicate, glucan, or other known binders. In another embodiment, the commercial package, feed additive, additive for the drinking water of the animal, or feed composition can further comprise inorganic / organic binders, essential oils, and / or organic acids. Any of the binder or carrier components can be exogenously added (i.e., not naturally present in combination with the Bacillus strains) and any of the compositions described herein can be present in dietary nutrient compositions or nutrient compositions (e.g., probiotic compositions).

[0187] In one aspect, any of the compositions described herein can further comprise an exogenously added (i.e., not naturally present with the Bacillus strain) nutrient component selected from the group consisting of a vitamin, an antibiotic, an enzyme, a water-soluble or water-insoluble monosaccharide, disaccharide, or polysaccharide, a fat, phosphorous, sodium bicarbonate, limestone, calcium, sodium, sulfur, magnesium, potassium, copper, iron, manganese, zinc, fish oil, raw seed, an antioxidant, and a starch. In one embodiment, the exogenously added nutrient component can be an enzyme and the enzyme can be selected from the group consisting of a galactosidase, a protease, a lipase, an amylase, a hemicellulase, an arabinoxylanase, a xylanase, a cellulase, an NSPase, a phytase, and combinations thereof.

[0188] The following examples are for illustrative purposes only. The examples are non- limiting, and are not intended to limit the invention in any way.EXAMPLE 1EFFECT OF BACILLUS STRAINS ON THE OF GROWTH OF FUSOBACTERIUM AND TRUEPERELLA

[0189] The objective of the assay to determine the effect of Bacillus strain 86 (NRRL No. B-50944), Bacillus strain 300 (NRRL No. B-50943), Bacillus strain 101 (NRRL No. B- 67218), Bacillus strain 102 (NRRL No. B-67276), Bacillus strain 177 (NRRL B-67275), and Bacillus strain 235 (NRRL B-67219) on the inhibition of growth of isolates of two subspecies of Fusobacterium necrophorum and isolates of Trueperella pyogenes from liver abscesses in feedlot cattle.Design and Methods

[0190] Eight isolates of Fusobacterium necrophorum subspecies necrophorum and subspecies funduliforme and five isolates of Trueperella pyogenes from liver abscesses in feedlot cattle were obtained from the collection of Dr. T.G. Nagaraja, Diagnostic Medicine and Pathobiology, Kansas State University. Isolates were tested against a panel of Bacillus strains (see Tables 1 and 2) using the cross-streak method on tryptic soy agar with 5% sheep’s blood. For the cross-streak method, tryptic soy agar plates with 5% sheep’s blood were streaked across the midline with a 1 pl calibrated loop of Bacillus culture and incubated for 24 hours at 37°C, then placed in sealed containers with GasPak anaerobic sachets to equilibrate for 4 hours. Working in an anaerobic chamber, Fusobacterium isolates were inoculated onto duplicate cross-streak plates with a 1 pl calibrated loop perpendicular to the Bacillus streak. After incubating in anaerobic jars at 37°C for 24 hours, Fusobacterium growth was examined, and a zone of inhibition was measured in mm from the edge of the Bacillus streak. Trueperella pyogenes isolates were tested with the same cross-streak method on the same media under aerobic growth conditions.Table 1. Bacillus isolate effect on growth inhibition of Fusobacterium isolates associated with bovine liver abscess.Table 2. Bacillus isolate effect on growth inhibition of Trueperella pyogenes isolates associated with bovine liver abscess.Bacillus strain 86 (NRRL No. B-50944), Bacillus strain 300 (NRRL No. B-50943), Bacillus strain 101 (NRRL No. B-67218), Bacillus strain 102 (NRRL No. B-67276), Bacillus strain 177 (NRRL B-67275), and Bacillus strain 235 (NRRL B-67219) show inhibitory activity against Fusobacterium isolates of bovine liver abscess origin, with two strains, Bacillus strains, 102 (B- 67276) and 177 (B-67275), showing very strong inhibition. Further Bacillus strain 86 (NRRLNo. B-50944), Bacillus strain 300 (NRRL No. B-50943), Bacillus strain 101 (NRRL No. B- 67218), Bacillus strain 102 (NRRL No. B-67276), Bacillus strain 177 (NRRL B-67275), and Bacillus strain 235 (NRRL B-67219) show inhibitory activity against Trueperella pyogenes isolates of bovine liver abscess origin.EXAMPLE 2EFFECT OF BACILLUS STRAINS ON THE OF GROWTH OF FUSOBACTERIUM AND TRUEPERELLA

[0191] The objective of the assay was to determine the effect of Bacillus strain 86 (NRRL No. B-50944), Bacillus strain 300 (NRRL No. B-50943), Bacillus strain 101 (NRRL No. B-67218), Bacillus strain 102 (NRRL No. B-67276), Bacillus strain 177 (NRRL B-67275), and Bacillus strain 235 (NRRL B-67219) on the inhibition of growth of isolates of Fusobacterium spp. obtained from ruminant abscesses.Design and Methods

[0192] Ten isolates of Fusobacterium necrophorum and one isolate of Fusobacterium varium of animal origin were obtained from Dr. Clifford Shipley, College of Veterinary Medicine, University of Illinois. Isolates were tested against a panel of Bacillus strains (see Table 3) using the cross-streak method on tryptic soy agar with 5% sheep’s blood. For the cross-streak method, tryptic soy agar plates with 5% sheep’s blood were streaked across the midline with a 1 pl calibrated loop of Bacillus culture and incubated for 24 hours at 37°C, then placed in sealed containers with GasPak anaerobic sachets to equilibrate for 4 hours. Working in an anaerobic chamber, Fusobacterium isolates were inoculated onto duplicate cross-streak plates with a 1 pl calibrated loop perpendicular to the Bacillus streak. After incubating in anaerobic jars at 37°C for 24 hours, Fusobacterium growth was examined, and a zone of inhibition was measured in mm from the edge of the Bacillus streak.Table 3. Bacillus isolate effect on growth inhibition of Fusobacterium spp. isolates associated with cervid, ovine and bovine disease.Bacillus strain 86 (NRRL No. B-50944), Bacillus strain 300 (NRRL No. B-50943), Bacillus strain 101 (NRRL No. B-67218), Bacillus strain 102 (NRRL No. B-67276), Bacillus strain 177 (NRRL B-67275), and Bacillus strain 235 (NRRL B-67219) show inhibitory activity against Fusobacterium isolates of cervid, ovine and bovine origin, with two strains, Bacillus strains, 102 (B-67276) and 177 (B-67275), showing very strong inhibition of Fusobacterium necrophorum and Fusobacterium varium in vitro.EXAMPLE 3RAPP (Randomly Amplified Polymorphic DNA) Analysis of DFM Strains

[0193] In order to obtain genetic barcodes or “fingerprints” of the Bacillus strains described herein, RAPD (randomly amplified polymorphic DNA) analysis was performed with each strain. Each isolate was cultured overnight in 10 ml of tryptic soy broth at 32°C.Thereafter, DNA was extracted from these cultures using Qiagen’s DNeasy Mini kit, followingthe protocol provided for Gram-positive bacteria. A GE Healthcare Illustra Ready-To Go RAPD kit was used to perform RAPD-PCR with each DNA sample to amplify genetic fragments of arbitrary length. RAPD primers that were used for RAPD-PCR analysis include the following: Primer 1 (5 -GGTGCGGGAA-3'; SEQ ID NO: 1) ; PRIMER 2 (5 - GTTTCGCTCC3'; SEQ ID NO: 2); and PRIMER 3 (5'-GTAGACCCGT-3'; SEQ ID NO: 3). PCR products were separated via gel electrophoresis, and banding patterns were detected and matched using BioNumerics software. As shown in Figs. 1 to 5, a common DNA fingerprint was not observed between the DFM strains. Therefore, Figs. 1 to 5 demonstrate that the DFM strains described herein are different strains, each having a unique DNA fingerprint.

Claims

WHAT IS CLAIMED IS:

1. A method of treating an animal wherein the animal has a disease caused by a Fusobacterium or Trueperella species, the method comprising the step of administering to the animal a feed composition or drinking water comprising an effective amount of an additive comprising an isolated Bacillus strain selected from the group consisting of Bacillus strain 86 (NRRL No. B-50944), Bacillus strain 300 (NRRL No. B-50943), Bacillus strain 101 (NRRL No. B-67218), Bacillus strain 235 (NRRL No. B-67219), Bacillus strain 177 (NRRL No. B- 67275), Bacillus strain 102 (NRRL No. B-67276), a strain having all of the idenmtifying characteristics of Bacillus strain 86 (NRRL No. B-50944), a strain having all of the identifying characteristics of Bacillus strain 300 (NRRL No. B-50943), a strain having all of the identifying characteristics of Bacillus strain 101 (NRRL No. B-67218), a strain having all of the identifying characteristics of Bacillus strain 235 (NRRL No. B-67219), a strain having all of the identifying characteristics of Bacillus strain 177 (NRRL No. B-67275), a strain having all of the identifying characteristics of Bacillus strain 102 (NRRL No. B-67276), and combinations thereof, wherein the Bacillus strain improves the disease caused by the Fusobacterium or Trueperella species.

2. The method of claim 1 wherein the animal is selected from the group consisting of a poultry species, a porcine species, a bovine species, an ovine species, a cervid species, an equine species, and a companion animal.

3. The method of claim 1 or 2 wherein the animal is a dairy animal.

4. The method of claim 3 wherein the dairy animal is selected from the group consisting of a cow, a goat, a sheep, and a buffalo.

5. The method of claim 1 or 2 wherein the animal is a deer.

6. The method of any one of claims 1 to 5 wherein the Bacillus strain produces an enzyme selected from the group consisting of an a-galactosidase, a protease, a lipase, an amylase, a xylanase, a cellulase, and combinations thereof.

7. The method of any one of claims 1 to 6 further comprising the step of administering to the animal another bacterial strain selected from the group consisting of another Bacillus strain, a lactic acid bacterial strain, and combinations thereof.

8. The method of any one of claims 1 to 7 wherein Bacillus strain 86 (NRRL No. B-50944), Bacillus strain 300 (NRRL No. B-50943), Bacillus strain 101 (NRRL No. B-67218), Bacillus strain 235 (NRRL No. B-67219), Bacillus strain 177 (NRRL No. B-67275), and Bacillus strain 102 (NRRL No. B-67276) are all administered to the animal.

9. The method of any one of claims 1 to 7 wherein the strain administered isBacillus strain 101 (NRRL No. B-67218).

10. The method of any one of claims 1 to 7 wherein the strain administered is Bacillus strain 235 (NRRL No. B-67219).1 1 . The method of any one of claims 1 to 7 wherein the strain administered is Bacillus strain 86 (NRRL No. B-50944).

12. The method of any one of claims 1 to 7 wherein the strain administered is Bacillus strain 300 (NRRL No. B-50943).

13. The method of any one of claims 1 to 7 wherein the strain administered is Bacillus strain 177 (NRRL No. B-67275).

14. The method of any one of claims 1 to 7 wherein the strain administered is Bacillus strain 102 (NRRL No. B-67276).

15. The method of any one of claims 1 to 7 wherein the strains administered are Bacillus strains 102 (NRRL No. B-67276) and 177 (NRRL No. B-67275).

16. The method of any one of claims 1 to 15 wherein the disease is caused by a Fusobacterium species.

17. The method of claim 16 wherein the Fusobacterium species is selected from the group consisting of Fusobacterium varium, Fusobacterium necrophorum, and combinations thereof.

18. The method of claim 16 wherein the Fusobacterium species is a Fusoobacterium subspecies selected from the group consisting of Fusobacterium necrophorum necrophorum, Fusobacterium necrophorum funduliforme, and combinations thereof.

19. The method of any one of claims 1 to 15 wherein the disease is caused by a Trueperella species.

20. The method of claim 19 wherein the Trueperella species is Trueperella pyogenes.

21. The method of any one of claims 1 to 20 wherein the disease is selected from the group consisting of liver abscess, necrobacillosis, necrotic laryngitis, lameness, mastitis, metritis, pyometra, pneumonia, and combinations thereof.

22. The method of claim 21 wherein the necrobacillosis is selected from bull nose disease and foot rot.

23. The method of claim 21 wherein the necrotic laryngitis is calf diphtheria.

24. The method of any one of claims 1 to 23 wherein the improvement in the disease caused by the Fusobacterium or Trueperella species results in an improvement selected from the group consisting of an improvement in the health of the animal, an improvement incarcass quality, an improvement in production efficiency, an improvement in condemnation rate, an improvement in feed efficiency, and combinations thereof.

25. The method of any one of claims 1 to 24 wherein each of the Bacillus strains is administered in the feed composition at a dose of about 1.0 x 103CFU / pound of the feed composition to about 5.0 x 1012CFU / pound of the feed composition.

26. The method of any one of claims 1 to 24 wherein each of the Bacillus strains is administered in the feed composition at a dose of about 0.5 x 107CFU / pound of the feed composition to about 5.0 x 107CFU / pound of the feed composition.

27. The method of any one of claims 1 to 24 wherein each of the Bacillus strains is administered in the feed composition at a dose greater than about 5.0 x 106CFU / pound of the feed composition.

28. The method of any one of claims 1 to 27 further comprising the step of administering an antibiotic to the animal.

29. The method of any one of claims 1 to 28 further comprising the step of administering to the animal an enzyme selected from the group consisting of a galactosidase, a protease, a lipase, an amylase, a hemicellulase, an arabinoxylanase, a xylanase, a cellulase, an NSPase, a phytase, and combinations thereof.

30. The method of claim 29 wherein the enzyme is an NSPase or a phytase.

31. The method of any one of claims 1 to 30 wherein the feed composition is administered daily to the animal.

32. A method of treating an animal wherein the animal has a disease selected from the group consisting of liver abscess, necrotic laryngitis, calf diphtheria, mastitis, pyometra, and bull nose disease, the method comprising the step of administering to the animal a feed composition or drinking water comprising an effective amount of an additive comprising an isolated Bacillus strain selected from the group consisting of Bacillus strain 86 (NRRL No. B- 50944), Bacillus strain 300 (NRRL No. B-50943), Bacillus strain 101 (NRRL No. B-67218), Bacillus strain 235 (NRRL No. B-67219), Bacillus strain 177 (NRRL No. B-67275), Bacillus strain 102 (NRRL No. B-67276), a strain having all of the identifying characteristics of Bacillus strain 86 (NRRL No. B-50944), a strain having all of the identifying characteristics of Bacillus strain 300 (NRRL No. B-50943), a strain having all of the identifying characteristics of Bacillus strain 101 (NRRL No. B-67218), a strain having all of the identifying characteristics of Bacillus strain 235 (NRRL No. B-67219), a strain having all of the identifying characteristics of Bacillus strain 177 (NRRL No. B-67275), a strain having all of the identifying characteristics of Bacillus strain 102 (NRRL No. B-67276), and combinations thereof, wherein the Bacillus strainimproves the disease selected from the group consisting of liver abscess, necrotic laryngitis, calf diphtheria, mastitis, pyometra, and bull nose disease.

33. The method of claim 32 wherein the animal is selected from the group consisting of a poultry species, a porcine species, a bovine species, an ovine species, a cervid species, an equine species, and a companion animal.

34. The method of claim 32 or 33 wherein the animal is a dairy animal.

35. The method of claim 34 wherein the dairy animal is selected from the group consisting of a cow, a goat, a sheep, and a buffalo.

36. The method of claim 32 or 33 wherein the animal is a deer.

37. The method of any one of claims 32 to 36 wherein the Bacillus strain produces an enzyme selected from the group consisting of an a-galactosidase, a protease, a lipase, an amylase, a xylanase, a cellulase, and combinations thereof.

38. The method of any one of claims 32 to 37 further comprising the step of administering to the animal another bacterial strain selected from the group consisting of another Bacillus strain, a lactic acid bacterial strain, and combinations thereof.

39. The method of any one of claims 32 to 38 wherein Bacillus strain 86 (NRRL No. B-50944), Bacillus strain 300 (NRRL No. B-50943), Bacillus strain 101 (NRRL No. B-67218), Bacillus strain 235 (NRRL No. B-67219), Bacillus strain 177 (NRRL No. B-67275), and Bacillus strain 102 (NRRL No. B-67276) are all administered to the animal.

40. The method of any one of claims 32 to 38 wherein the strain administered is Bacillus strain 101 (NRRL No. B-67218).

41. The method of any one of claims 32 to 38 wherein the strain administered is Bacillus strain 235 (NRRL No. B-67219).

42. The method of any one of claims 32 to 38 wherein the strain administered is Bacillus strain 86 (NRRL No. B-50944).

43. The method of any one of claims 32 to 38 wherein the strain administered is Bacillus strain 300 (NRRL No. B-50943).

44. The method of any one of claims 32 to 38 wherein the strain administered is Bacillus strain 177 (NRRL No. B-67275).

45. The method of any one of claims 32 to 38 wherein the strain administered is Bacillus strain 102 (NRRL No. B-67276).

46. The method of any one of claims 32 to 38 wherein the strains administered areBacillus strains 102 (NRRL No. B-67276) and 177 (NRRL No. B-67275).

47. The method of any one of claims 32 to 46 wherein the disease is liver abscess.

48. The method of any one of claims 32 to 46 wherein the disease is necrotic laryngitis.

49. The method of any one of claims 32 to 48 wherein the improvement in the disease results in an improvement selected from the group consisting of an improvement in health of the animal, an improvement in carcass quality, an improvement in production efficiency, an improvement in condemnation rate, an improvement in feed efficiency, and combinations thereof.

50. The method of any one of claims 32 to 49 wherein each of the Bacillus strains is administered in the feed composition at a dose of about 1.0 x 103CFU / pound of the feed composition to about 5.0 x 1012CFU / pound of the feed composition.

51. The method of any one of claims 32 to 49 wherein each of the Bacillus strains is administered in the feed composition at a dose of about 0.5 x 107CFU / pound of the feed composition to about 5.0 x 107CFU / pound of the feed composition.

52. The method of any one of claims 32 to 49 wherein each of the Bacillus strains is administered in the feed composition at a dose greater than about 5.0 x 106CFU / pound of the feed composition.

53. The method of any one of claims 32 to 52 further comprising the step of administering an antibiotic to the animal.

54. The method of any one of claims 32 to 53 further comprising the step of administering to the animal an enzyme selected from the group consisting of a galactosidase, a protease, a lipase, an amylase, a hemicellulase, an arabinoxylanase, a xylanase, a cellulase, an NSPase, a phytase, and combinations thereof.

55. The method of claim 54 wherein the enzyme is an NSPase or a phytase.

56. The method of any one of claims 32 to 55 wherein the feed composition is administered daily to the animal.

57. The method of any one of claims 1 to 56 wherein the additive is added to the feed composition or drinking water in the form of a concentrate.

58. The method of any one of claims 1 to 56 wherein the additive is added to the feed composition or the drinking water in the form of a superconcentrate.

59. The method of any one of claims 1 to 58 wherein the additive is added to the feed composition or the drinking water in dry form.

60. The method of any one of claims 1 to 59 wherein the feed composition is in pelleted form.

61. The method of any one of claims 1 to 60 wherein the strains are in a form selected from the group consisting of a powder, a liquid, and a pellet form.

62. The method of any one of claims 1 to 61 wherein the additive further comprises a carrier for the Bacillus strains.

63. The method of claim 62 wherein the carrier is selected from the group consisting of a bran, rice hulls, a salt, a dextrin, and combinations thereof.

64. The method of claim 61 wherein the strains are in powder form.

65. The method of claim 61 wherein the strains are in liquid form.

66. The method of any one of claims 1 to 65 wherein the additive further comprises a binder.

67. The method of claim 66 wherein the binder is selected from the group consisting of clay, yeast cell wall components, aluminum silicate, glucan, and combinations thereof.

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