Microorganisms for animals

Abstract

The present invention relates to direct-fed microorganisms for improving the reproductive performance of animals, reducing death losses and / or fasting events in animals, improving milk production and / or milk quality in dairy animals, and inhibiting diseases or disorders in dairy animals selected from the group consisting of: milk fever, ketosis, retained placenta, metritis, pneumonia, and ectopic abomasum. More specifically, the present invention relates to isolated Bacillus strains (86, 300, 101, 235, 177, and 102) and strains having all the identifying characteristics of these strains for use including the above uses.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims priority under 35 U.S.C. 119(e) to U.S. Provisional Application Serial No. 62 / 740,169, filed on October 2, 2018, the entire disclosure of which is incorporated herein by reference. Technical Field

[0003] The present invention relates to direct-fed microorganisms for improving the reproductive performance of animals, reducing death losses and / or off-feed events in animals, improving milk production and / or milk quality in dairy animals, and inhibiting diseases or disorders in dairy animals selected from the group consisting of: milk fever, ketosis, retained placenta, metritis, pneumonia, and ectopic abomasum. More specifically, the present invention relates to isolated Bacillus sp. Bacillus ) strains 86, 300, 101, 235, 177 and 102, and strains having all the identifying characteristics of these strains, for uses including the above uses.

[0004] Background and Summary of the Invention

[0005] The present invention relates to direct-fed microbial (DFM) compositions. Reproductive performance is very important in all livestock production. In dairy cows, timely breeding of cows is critical to ensure that the cows reach their potential in lifetime milk production. Delayed conception, high abortion rates, and low pregnancy rates can have a negative impact on the overall milk production of the herd, and cows with reproductive difficulties often need to be removed from the herd. Improvements in reproductive performance can create value for the dairy industry in a variety of ways: through increased milk production, through additional calves born, and through savings in replacement cow costs. In addition, improvements in reproduction save costs associated with additional labor and technicians to perform inseminations, therapeutic drugs, and labor and costs for synchronization of insemination or ovulation times.

[0006] Likewise, a significant financial and labor investment is required in raising livestock. The costs of feed, fresh water, shelter, vaccination and medical treatment, handling and milking are several of the significant expenses associated with managing livestock, including dairy cows. For example, the cost of raising a heifer replacement animal can exceed $1,500. In the dairy industry, controlling cow mortality is critical from both an economic and dairy animal welfare perspective. In recent decades, adult cow mortality has been increasing and includes death losses due to calving disorders, digestive disorders, motility disorders, metabolic disorders, udder / teater disorders, accidents, and unknown causes. New technologies are needed in the dairy industry and the livestock industry as a whole to reduce mortality loss rates.

[0007] As the world population increases, the demand for high-quality protein products, including dairy products, continues to grow, but pressure to reduce the environmental impact of agriculture by reducing the use of land and water resources is also increasing. Improvements in total milk production per cow and milk components, including fat and protein, can help meet both needs, thereby meeting the growing demand for higher milk production without additional animal resources. In addition, efficient milk production is necessary for the economic viability of the dairy industry. Improving the fat and protein percentages in milk results in higher quality milk, which in turn leads to greater profits for the dairy industry.

[0008] Applicants have developed DFM compositions that result in improved reproductive performance of animals, reduced mortality losses and / or fasting events in animals, improved milk production and / or quality in dairy animals, and inhibition of diseases or disorders in dairy animals, wherein the diseases or disorders are selected from the group consisting of milk fever, ketosis, retained placenta, metritis, pneumonia, and ectopic abomasum. The DFM compositions described herein provide commercial benefits by providing all of these properties or a combination thereof.

[0009] Provided are methods and compositions for improving the reproductive performance of animals, reducing the death loss and / or fasting events of animals, improving the milk production and / or milk quality of dairy animals, and inhibiting diseases or disorders selected from the following in dairy animals: lactogenic fever, ketosis, retained placenta, metritis, pneumonia, and ectopic abomasum. In various embodiments, the animal can be selected from poultry species, porcine species, bovine species, ovine species, equine species, and companion animals. In embodiments where the animal is a porcine species, the porcine species can be selected from growth-finished pigs, nursery pigs, sows, and breeding pigs. In another embodiment, the animal can be a dairy animal selected from cows, goats, sheep, or buffaloes.

[0010] In various embodiments, the composition used in the methods described herein can be a commercial package, a feed additive for an animal feed composition, an additive for animal drinking water, or an animal feed composition (e.g., a complete feed), each comprising all of the isolated strains, including 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).

[0011] In another embodiment, a method for improving the reproductive performance of an animal is provided. The method comprises the step of administering to the animal a feed composition or drinking water comprising an effective amount of an additive, wherein the additive comprises 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 the identifying characteristics of Bacillus strain 86 (NRRL No. B-50944), a strain having all the identifying characteristics of Bacillus strain 300 (NRRL No. B-50943), a strain having all the identifying characteristics of Bacillus strain 101 (NRRL No. B-67218), a strain having all the identifying characteristics of Bacillus strain 235 (NRRL No. B-67219), a strain having all the identifying characteristics of Bacillus strain 177 (NRRL No. B-67275), a strain having all the identifying characteristics of Bacillus strain 102 (NRRL No. B-67276), a strain having all the identifying characteristics of Bacillus strain 86 (NRRL No. B-50944), a strain having all the identifying characteristics of Bacillus strain 300 (NRRL No. B-50943), a strain having all the identifying characteristics of Bacillus strain 101 (NRRL No. B-67218), a strain having all the identifying characteristics of Bacillus strain 235 (NRRL No. B-67219), a strain having all the identifying characteristics of Bacillus strain 177 (NRRL No. B-67275), a strain having all the identifying characteristics of Bacillus strain 102 (NRRL No. B-67276), and combinations thereof, wherein the Bacillus strain improves the reproductive performance of the animal.

[0012] In another embodiment, a method of reducing mortality losses in an animal and / or reducing fasting events in an animal is provided. The method comprises the step of administering to the animal a feed composition or drinking water comprising an effective amount of an additive, wherein the additive comprises 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 the identifying characteristics of Bacillus strain 86 (NRRL No. B-50944), a strain having all the identifying characteristics of Bacillus strain 300 (NRRL No. B-50943), a strain having all the identifying characteristics of Bacillus strain 101 (NRRL No. B-67218), a strain having all the identifying characteristics of Bacillus strain 235 (NRRL No. B-67219), (NRRL No. B-67219), strains having all the identifying characteristics of Bacillus strain 177 (NRRL No. B-67275), strains having all the identifying characteristics of Bacillus strain 102 (NRRL No. B-67276), and combinations thereof, wherein the Bacillus strains reduce mortality losses in animals and / or reduce fasting episodes in animals.

[0013] In another embodiment, a method of improving milk production and / or milk quality in a dairy animal is provided. The method comprises the step of administering to the animal a feed composition or drinking water comprising an effective amount of an additive, wherein the additive comprises 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 the identifying characteristics of Bacillus strain 86 (NRRL No. B-50944), a strain having all the identifying characteristics of Bacillus strain 300 (NRRL No. B-50943), a strain having all the identifying characteristics of Bacillus strain 101 (NRRL No. B-67218), a strain having all the identifying characteristics of Bacillus strain 235 (NRRL No. B-67219), (NRRL No. B-67219), a strain having all the identifying characteristics of Bacillus strain 177 (NRRL No. B-67275), a strain having all the identifying characteristics of Bacillus strain 102 (NRRL No. B-67276), and combinations thereof, wherein the Bacillus strain improves milk production and / or milk quality in the animal.

[0014] In another exemplary embodiment, a method for inhibiting a disease or disorder in a milk-producing animal is provided, wherein the disease or disorder is selected from the group consisting of milk fever, ketosis, retained placenta, metritis, pneumonia, and ectopic abomasum. When the disease is pneumonia, the method can also be used for other types of animals. The method comprises the step of administering to the animal a feed composition or drinking water comprising an effective amount of an additive, wherein the additive comprises 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 the identifying characteristics of Bacillus strain 86 (NRRL No. B-50944), a strain having all the identifying characteristics of Bacillus strain 300 (NRRL No. B-50943), a strain having all the identifying characteristics of Bacillus strain 101 (NRRL No. B-67218), a strain having all the identifying characteristics of Bacillus strain 235 (NRRL No. B-67219), (NRRL No. B-67219), a strain having all the identifying characteristics of Bacillus strain 177 (NRRL No. B-67275), a strain having all the identifying characteristics of Bacillus strain 102 (NRRL No. B-67276), and combinations thereof, wherein the Bacillus strain inhibits the disease or disorder, wherein the disease or disorder is selected from the group consisting of milk fever, ketosis, retained placenta, metritis, pneumonia, and ectopic abomasum.

[0015] The following clauses and combinations thereof provide various additional exemplary aspects of the invention described herein. Various embodiments described in any other section of this patent application (including sections entitled "Detailed Description" and Examples) are applicable to any of the following embodiments of the invention described in the following numbered clauses.

[0016] 1. A method for improving the reproductive performance of an animal, the method comprising the step of administering to the animal a feed composition or drinking water comprising an effective amount of an additive, the 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 the identifying characteristics of Bacillus strain 86 (NRRL No. B-50944), a strain having all the identifying characteristics of Bacillus strain 300 (NRRL No. B-50943), a strain having all the identifying characteristics of Bacillus strain 101 (NRRL No. B-67218), a strain having all the identifying characteristics of Bacillus strain 235 (NRRL No. B-67219), a strain having all the identifying characteristics of Bacillus strain 177 (NRRL No. B-67275), a strain having all the identifying characteristics of Bacillus strain 102 (NRRL No. B-67276), and combinations thereof, wherein the Bacillus strain improves the reproductive performance of the animal.

[0017] 2. A method of reducing mortality losses and / or reducing fasting events in animals, the method comprising the step of administering to the animal a feed composition or drinking water comprising an effective amount of an additive, the 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 the identifying characteristics of Bacillus strain 86 (NRRL No. B-50944), a strain having all the identifying characteristics of Bacillus strain 300 (NRRL No. B-50943), a strain having all the identifying characteristics of Bacillus strain 101 (NRRL No. B-67218), a strain having all the identifying characteristics of Bacillus strain 235 (NRRL No. B-67219), a strain having all the identifying characteristics of Bacillus strain 177 (NRRL No. B-67275), a strain having all the identifying characteristics of Bacillus strain 102 (NRRL No. B-67276), and combinations thereof, wherein the Bacillus strain reduces mortality losses in animals and / or reduces fasting episodes in animals.

[0018] 3. The method of clause 1 or 2, wherein the animal is selected from poultry species, porcine species, bovine species, ovine species, equine species, and companion animals.

[0019] 4. The method of any one of clauses 1 to 3, wherein the animal is a porcine species and the porcine species is selected from the group consisting of finishing pigs, nursery pigs, sows and breeder pigs.

[0020] 5. The method of clause 1 or 2, wherein the animal is a milk-producing animal.

[0021] 6. The method of clause 5, wherein the milk-producing animal is selected from a cow, a goat, a sheep or a buffalo.

[0022] 7. The method of any one of clauses 1 to 6, wherein the Bacillus strain produces an enzyme selected from the group consisting of α-galactosidase, protease, lipase, amylase, xylanase, cellulase, and combinations thereof.

[0023] 8. The method of any one of clauses 1 to 7, further comprising the step of administering to the animal another bacterial strain selected from another Bacillus strain, a lactic acid bacteria strain, and combinations thereof.

[0024] 9. The method of any of clauses 1-8, 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.

[0025] 10. The method of any one of clauses 1 to 8, wherein the strain applied is Bacillus strain 101 (NRRL No. B-67218) or a strain having all the identifying characteristics of Bacillus strain 101 (NRRL No. B-67218).

[0026] 11. The method of any one of clauses 1 to 8, wherein the strain applied is Bacillus strain 235 (NRRL No. B-67219) or a strain having all of the identifying characteristics of Bacillus strain 235 (NRRL No. B-67219).

[0027] 12. The method of any one of clauses 1 to 8, wherein the strain applied is Bacillus strain 86 (NRRL No. B-50944) or a strain having all of the identifying characteristics of Bacillus strain 86 (NRRL No. B-50944).

[0028] 13. The method of any one of clauses 1 to 8, wherein the strain applied is Bacillus strain 300 (NRRL No. B-50943) or a strain having all of the identifying characteristics of Bacillus strain 300 (NRRL No. B-50943).

[0029] 14. The method of any one of clauses 1 to 8, wherein the strain applied is Bacillus strain 177 (NRRL No. B-67275) or a strain having all of the identifying characteristics of Bacillus strain 177 (NRRL No. B-67275).

[0030] 15. The method of any one of clauses 1 to 8, wherein the strain applied is Bacillus strain 102 (NRRL No. B-67276) or a strain having all of the identifying characteristics of Bacillus strain 102 (NRRL No. B-67276).

[0031] 16. The method of any one of clauses 1 to 8, wherein the strain applied is Bacillus strain 101 (NRRL No. B-67218).

[0032] 17. The method of any one of clauses 1 to 8, wherein the strain applied is Bacillus strain 235 (NRRL No. B-67219).

[0033] 18. The method of any one of clauses 1 to 8, wherein the strain applied is Bacillus strain 86 (NRRL No. B-50944).

[0034] 19. The method of any one of clauses 1 to 8, wherein the strain applied is Bacillus strain 300 (NRRL No. B-50943).

[0035] 20. The method of any one of clauses 1 to 8, wherein the strain applied is Bacillus strain 177 (NRRL No. B-67275).

[0036] 21. The method of any one of clauses 1 to 8, wherein the strain applied is Bacillus strain 102 (NRRL No. B-67276).

[0037] 22. The method of any one of clauses 1 to 21, wherein the 3 CFU / lb of feed composition to approximately 5.0 x 10 12 Dosage of CFU / lb of feed composition Each Bacillus strain was administered in the feed composition.

[0038] 23. The method of any one of clauses 1 to 21, wherein the7 CFU / lb of feed composition to approximately 5.0 x 10 7 Dosage of CFU / lb of feed composition Each Bacillus strain was administered in the feed composition.

[0039] 24. The method of any one of clauses 1 to 21, wherein the 6 Dosage of CFU / lb of feed composition Each Bacillus strain was administered in the feed composition.

[0040] 25. The method of any one of clauses 1 to 24 further comprising the step of administering an antibiotic to the animal.

[0041] 26. The method of clause 25, wherein the antibiotic is selected from erythromycin, levofloxacin, trimethoprim / sulfamethoxazole, trimethoprim, daptomycin, rifampicin, tylosin, tilmicosin, vancomycin, monensin, lasalocid, and lelomycin propionate.

[0042] 27. The method of any one of clauses 1 to 26, further comprising the step of administering to the animal an enzyme selected from the group consisting of β-galactosidase, protease, lipase, amylase, hemicellulase, arabinoxylanase, xylanase, cellulase, NSPase, phytase, and combinations thereof.

[0043] 28. The method of clause 27, wherein the enzyme is NSPase or phytase.

[0044] 29. The method of clause 1 or 2, wherein the animal is a cow and the Bacillus strain is administered during lactation.

[0045] 30. The method of clause 1 or 2, wherein the animal is a cow and the Bacillus strain is administered during gestation.

[0046] 31. The method of any one of clauses 1 to 30, wherein the feed composition is administered to the animal daily.

[0047] 32. The method of any one of clauses 1 or 3-31 wherein the improvement in reproductive performance is selected from a reduction in the number of conception services, an increase in estrus detection, an increase in conception rate, a reduction in miscarriage and an increase in pregnancy rate.

[0048] 33. A method of improving milk production and / or milk quality in a dairy animal, the method comprising the step of administering to the animal a feed composition or drinking water comprising an effective amount of an additive, the 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 the identifying characteristics of Bacillus strain 86 (NRRL No. B-50944), a strain having all the identifying characteristics of Bacillus strain 300 (NRRL No. B-50943), a strain having Bacillus strain 101 (NRRL No. B-67218), (NRRL No. B-67218), a strain having all the identifying characteristics of Bacillus strain 235 (NRRL No. B-67219), a strain having all the identifying characteristics of Bacillus strain 177 (NRRL No. B-67275), a strain having all the identifying characteristics of Bacillus strain 102 (NRRL No. B-67276), and combinations thereof, wherein the Bacillus strain improves milk production and / or milk quality in the animal.

[0049] 34. A method of inhibiting a disease or disorder in a milk-producing animal, wherein the disease or disorder is selected from the group consisting of milk fever, ketosis, retained placenta, metritis, pneumonia, and ectopic abomasum, 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 the identifying characteristics of Bacillus strain 86 (NRRL No. B-50944), a strain having all the identifying characteristics of Bacillus strain 300 (NRRL No. B-50943), a strain having Bacillus strain 101 (NRRL No. B-67218), (NRRL No. B-67218), a strain having all the identifying characteristics of Bacillus strain 235 (NRRL No. B-67219), a strain having all the identifying characteristics of Bacillus strain 177 (NRRL No. B-67275), a strain having all the identifying characteristics of Bacillus strain 102 (NRRL No. B-67276), and combinations thereof, wherein the Bacillus strain inhibits a disease or disorder in the milk producing animal, wherein the disease or disorder is selected from the group consisting of milk fever, ketosis, retained placenta, metritis, pneumonia, and ectopic abomasum.

[0050] 35. The method of clause 33 or 34, wherein the milk producing animal is selected from a cow, goat, sheep or buffalo.

[0051] 36. The method of any one of clauses 33 to 35, wherein the Bacillus strain produces an enzyme selected from the group consisting of alpha-galactosidase, protease, lipase, amylase, xylanase, cellulase, and combinations thereof.

[0052] 37. The method of any one of clauses 33 to 36 further comprising the step of administering to the animal another bacterial strain selected from another Bacillus strain, a lactic acid bacteria strain, and combinations thereof.

[0053] 38. The method of any of clauses 33-37, 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.

[0054] 39. The method of any of clauses 33-37, wherein the strain applied is Bacillus strain 101 (NRRL No. B-67218) or a strain having all of the identifying characteristics of Bacillus strain 101 (NRRL No. B-67218).

[0055] 40. The method of any of clauses 33-37, wherein the strain administered is Bacillus strain 235 (NRRL No. B-67219) or a strain having all of the identifying characteristics of Bacillus strain 235 (NRRL No. B-67219).

[0056] 41. The method of any of clauses 33-37, wherein the strain administered is Bacillus strain 86 (NRRL No. B-50944) or a strain having all of the identifying characteristics of Bacillus strain 86 (NRRL No. B-50944).

[0057] 42. The method of any of clauses 33-37, wherein the strain administered is Bacillus strain 300 (NRRL No. B-50943) or a strain having all of the identifying characteristics of Bacillus strain 300 (NRRL No. B-50943).

[0058] 43. The method of any of clauses 33-37, wherein the strain administered is Bacillus strain 177 (NRRL No. B-67275) or a strain having all of the identifying characteristics of Bacillus strain 177 (NRRL No. B-67275).

[0059] 44. The method of any of clauses 33-37, wherein the strain administered is Bacillus strain 102 (NRRL No. B-67276) or a strain having all of the identifying characteristics of Bacillus strain 102 (NRRL No. B-67276).

[0060] 45. The method of any of clauses 33-37, wherein the strain applied is Bacillus strain 101 (NRRL No. B-67218).

[0061] 46. ​​The method of any of clauses 33-37, wherein the strain applied is Bacillus strain 235 (NRRL No. B-67219).

[0062] 47. The method of any of clauses 33-37, wherein the strain applied is Bacillus strain 86 (NRRL No. B-50944).

[0063] 48. The method of any of clauses 33-37, wherein the strain applied is Bacillus strain 300 (NRRL No. B-50943).

[0064] 49. The method of any of clauses 33-37, wherein the strain applied is Bacillus strain 177 (NRRL No. B-67275).

[0065] 50. The method of any of clauses 33-37, wherein the strain applied is Bacillus strain 102 (NRRL No. B-67276).

[0066] 51. The method of any one of clauses 33 to 50, wherein the 3 CFU / lb of feed composition to approximately 5.0 x 10 12 Dosage of CFU / lb of feed composition Each Bacillus strain was administered in the feed composition.

[0067] 52. The method of any one of clauses 33 to 50, wherein the 7 CFU / lb of feed composition to approximately 5.0 x 10 7 Dosage of CFU / lb of feed composition Each Bacillus strain was administered in the feed composition.

[0068] 53. The method of any one of clauses 33 to 50, wherein the 6 Dosage of CFU / lb of feed composition Each Bacillus strain was administered in the feed composition.

[0069] 54. The method of any one of clauses 33-53, further comprising the step of administering an antibiotic to the animal.

[0070] 55. The method of clause 54, wherein the antibiotic is selected from erythromycin, levofloxacin, trimethoprim / sulfamethoxazole, trimethoprim, daptomycin, rifampicin, tylosin, tilmicosin, vancomycin, monensin, lasalocid, and lelomycin propionate.

[0071] 56. The method of any one of clauses 33 to 55, further comprising the step of administering to the animal an enzyme selected from the group consisting of β-galactosidase, protease, lipase, amylase, hemicellulase, arabinoxylanase, xylanase, cellulase, NSPase, phytase and combinations thereof.

[0072] 57. The method of clause 56, wherein the enzyme is NSPase or phytase.

[0073] 58. The method of any one of clauses 33 to 57, wherein the Bacillus strain is administered during lactation.

[0074] 59. The method of any one of clauses 33 to 57, wherein the Bacillus strain is administered during pregnancy.

[0075] 60. The method of any one of clauses 33 to 59, wherein the feed composition is administered to the animal daily.

[0076] 61. The method of any one of clauses 33 or 35-60 wherein the improvement in milk yield and / or milk quality is selected from: increasing milk yield, increasing milk fat percentage, increasing milk protein, increasing total fat corrected milk, increasing energy corrected milk yield and reducing somatic cell count.

[0077] 62. The method of any one of clauses 1, 2 or 5-61 wherein the animal is a cow.

[0078] 63. A commercial package for animal use comprising all isolated strains including 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).

[0079] 64. A feed additive for animal feed comprising all isolated strains, wherein the isolated strains include 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).

[0080] 65. An additive for animal drinking water comprising all isolated strains, wherein the isolated strains include 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).

[0081] 66. An animal feed composition comprising all isolated strains including 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).

[0082] 67. A commercial package, a feed additive, an additive for drinking water for an animal or an animal feed composition according to any one of clauses 63 to 66 for improving the reproductive performance of said animal.

[0083] 68. A commercial package, a feed additive, an additive for drinking water for animals or an animal feed composition as claimed in any one of clauses 63 to 66 for use in reducing mortality losses and / or fasting events in animals.

[0084] 69. A commercial package, a feed additive, an additive for drinking water for animals or an animal feed composition according to any of clauses 63 to 66 for improving milk production and / or milk quality in an animal.

[0085] 70. A commercial package, a feed additive, an additive for animal drinking water or an animal feed composition as claimed in any one of clauses 63 to 66 for use in inhibiting a disease or disorder in an animal selected from the group consisting of: lactogenic fever, ketosis, retained placenta, metritis, pneumonia and ectopic abomasum.

[0086] 71. A feed additive or an additive for drinking water for animals as defined in item 64 or 65 in the form of a concentrate.

[0087] 72. A feed additive or an additive for drinking water for animals as defined in item 64 or 65 in the form of a super concentrate.

[0088] 73. A feed additive, a feed composition or an additive for drinking water for animals according to any one of clauses 64 to 72 in dry form.

[0089] 74. The feed composition of clause 73 in the form of pellets.

[0090] 75. A commercial package, a feed additive or an additive for drinking water for animals according to any one of clauses 63-65 or 67-74, wherein the strain is in a form selected from the group consisting of a powder, a liquid and a pelleted form.

[0091] 76. A commercial package, a feed additive, an additive for drinking water for animals or a feed composition according to any one of clauses 63 to 75, further comprising a carrier of said Bacillus strain.

[0092] 77. The commercial package, feed additive, additive for animal drinking water or feed composition of clause 76, wherein the carrier is selected from the group consisting of bran, rice hulls, salt, dextrin and combinations thereof.

[0093] 78. A commercial package, a feed additive, an additive for drinking water for animals or a feed composition according to any of clauses 63 to 77 in a bag.

[0094] 79. The commercial bag, feed additive, additive for drinking water for animals or feed composition of clause 78, wherein the bag is a plastic bag.

[0095] 80. The commercial package, feed additive, additive for drinking water for animals or feed composition of any one of clauses 63 to 79, further comprising instructions for use of the one or more Bacillus strains.

[0096] 81. A commercial package, feed additive, feed composition, or additive for drinking water for animals of any of clauses 63 to 80 in a 20-pound bag.

[0097] 82. A commercial package, feed additive, feed composition, or additive for drinking water for animals of any of clauses 63 to 80 in a 50-pound bag.

[0098] 83. A feed additive or an additive for drinking water for animals according to any of clauses 64, 65, 67-73 or 75-82 in powder form.

[0099] 84. A feed additive or an additive for drinking water for animals as claimed in any one of clauses 64, 65, 67-72, 75 or 78-80 in liquid form.

[0100] 85. A commercial package, a feed additive, an additive for drinking water for animals or a feed composition as claimed in any one of clauses 63 to 84 in a container for commercial use.

[0101] 86. The commercial package, feed additive, additive for drinking water for animals or feed composition of clause 85, wherein the container comprises plastic.

[0102] 87. The commercial bag, feed additive, additive for drinking water for animals or feed composition of clause 85, wherein the container comprises paper.

[0103] 88. A commercial package, a feed additive, an additive for drinking water for animals or a feed composition according to any one of clauses 63 to 87, wherein the strain further comprises a binder.

[0104] 89. The commercial package, feed additive, additive for animal drinking water or feed composition of clause 88, wherein the binder is selected from clay, yeast cell wall components, aluminum silicate and dextran or a combination thereof. BRIEF DESCRIPTION OF THE DRAWINGS

[0105] Figure 1 The following figure shows (Figure A) total E. coli of enteropathogenic and enterohemorrhagic E. coli (16S)、 Heat-labile enterotoxin gene ( LT ), tight adhesion and elimination genes ( eae ) of the rectum, (Figure B) Clostridium perfringens ( Clostridium perfringens ) α and β toxin genes (respectively CPA and CPB ) and (C) during Bacillus supplementation with Aspergillus ( Aspergillus )( 18S ) genes (p ≤ 0.05).

[0106] Figure 2 Shown is a photograph of a gel demonstrating the RAPD PCR profiles (primers 1-6) of 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 2 consecutive lanes between the markers corresponds to primers 1-6 from left to right.

[0107] Figure 3 Shown is a photograph of a gel demonstrating the RAPD PCR characteristics of Bacillus strain 101 (primers 1-6). For primer 1, fuzzy 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 fuzzy band appears at approximately 400 base pairs. For primer 3, a fuzzy band appears at approximately 400 base pairs, fuzzy double bands appear at 600 and 700 base pairs, and fuzzy bands appear at 1000 and 1400 base pairs.

[0108] Figure 4Shown is a photograph of a gel demonstrating the RAPD PCR characteristics (primers 1-3) of Bacillus strain 102. For primer 1, an intense band appears at 900 bp, and fuzzy bands appear at 1200 and 1400 bp. For primer 2, intense bands appear at 600, 700, and 1000 bp. For primer 3, an intense band appears at 600 bp, double bands appear at 700 and 750 bp, double bands appear at 900 and 1100 bp, and fuzzy bands appear at 400 and greater than 1800 bp.

[0109] Figure 5 Shown is a photograph of a gel demonstrating the RAPD PCR characteristics (primers 2 and 3) of Bacillus strain 177. For primer 2, intense bands appear at 600, 700, and 1000 base pairs. For primer 3, an intense band appears at 600 base pairs, double bands appear at 700 and 750 base pairs, double bands appear at 900 and 1100 base pairs, and faint bands appear at 400 and greater than 1800 base pairs.

[0110] Figure 6 Shown is a photograph of a gel demonstrating the RAPD PCR characteristics (primers 1-6) of Bacillus strain 235. For Primer 1, fuzzy bands appear at approximately 1000 and greater than 1600 base pairs. For Primer 2, an intense band appears at 650 base pairs and a fuzzy band appears at approximately 400 base pairs. For Primer 3, fuzzy bands appear at approximately 400, 600, 1000, and 1400 base pairs. DETAILED DESCRIPTION

[0111] Provided are methods and compositions for improving the reproductive performance of animals, reducing the death loss and / or fasting events of animals, improving the milk production and / or milk quality of dairy animals, and inhibiting diseases or disorders selected from the following in dairy animals: milk fever, ketosis, retained placenta, metritis, pneumonia, and ectopic abomasum. Also provided are methods for inhibiting pneumonia in animals described herein other than dairy animals. In different embodiments, the animal can be selected from poultry species, pig species, cattle species, sheep species, horse species, and companion animals. In embodiments where the animal is a pig species, the pig species can be selected from growth-finished pigs, nursery pigs, sows, and breeding pigs. In another embodiment, the animal can be a dairy animal selected from cows, goats, sheep, or buffaloes.

[0112] In various embodiments, the composition used in the methods described herein can be a commercial package, a feed additive for an animal feed composition, an additive for animal drinking water, or an animal feed composition (e.g., a complete feed), each comprising all of the isolated strains, including 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).

[0113] In another embodiment, a method for improving the reproductive performance of an animal is provided. The method comprises the step of administering to the animal a feed composition or drinking water comprising an effective amount of an additive, wherein the additive comprises 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 the identifying characteristics of Bacillus strain 86 (NRRL No. B-50944), a strain having all the identifying characteristics of Bacillus strain 300 (NRRL No. B-50943), a strain having all the identifying characteristics of Bacillus strain 101 (NRRL No. B-67218), a strain having all the identifying characteristics of Bacillus strain 235 (NRRL No. B-67219), a strain having all the identifying characteristics of Bacillus strain 177 (NRRL No. B-67275), a strain having all the identifying characteristics of Bacillus strain 102 (NRRL No. B-67276), a strain having all the identifying characteristics of Bacillus strain 86 (NRRL No. B-50944), a strain having all the identifying characteristics of Bacillus strain 300 (NRRL No. B-50943), a strain having all the identifying characteristics of Bacillus strain 101 (NRRL No. B-67218), a strain having all the identifying characteristics of Bacillus strain 235 (NRRL No. B-67219), a strain having all the identifying characteristics of Bacillus strain 177 (NRRL No. B-67275), a strain having all the identifying characteristics of Bacillus strain 102 (NRRL No. B-67276), and combinations thereof, wherein the Bacillus strain improves the reproductive performance of the animal.

[0114] In yet another embodiment, a method of reducing mortality losses in an animal and / or reducing fasting events in an animal is provided. The method comprises the step of administering to the animal a feed composition or drinking water comprising an effective amount of an additive, wherein the additive comprises 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 the identifying characteristics of Bacillus strain 86 (NRRL No. B-50944), a strain having all the identifying characteristics of Bacillus strain 300 (NRRL No. B-50943), a strain having all the identifying characteristics of Bacillus strain 101 (NRRL No. B-67218), a strain having all the identifying characteristics of Bacillus strain 235 (NRRL No. B-67219), (NRRL No. B-67219), strains having all the identifying characteristics of Bacillus strain 177 (NRRL No. B-67275), strains having all the identifying characteristics of Bacillus strain 102 (NRRL No. B-67276), and combinations thereof, wherein the Bacillus strains reduce mortality losses in animals and / or reduce fasting episodes in animals.

[0115] In yet another embodiment, a method of improving milk production and / or milk quality in a dairy animal is provided. The method comprises the step of administering to the animal a feed composition or drinking water comprising an effective amount of an additive, wherein the additive comprises 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 the identifying characteristics of Bacillus strain 86 (NRRL No. B-50944), a strain having all the identifying characteristics of Bacillus strain 300 (NRRL No. B-50943), a strain having all the identifying characteristics of Bacillus strain 101 (NRRL No. B-67218), a strain having all the identifying characteristics of Bacillus strain 235 (NRRL No. B-67219), (NRRL No. B-67219), a strain having all the identifying characteristics of Bacillus strain 177 (NRRL No. B-67275), a strain having all the identifying characteristics of Bacillus strain 102 (NRRL No. B-67276), and combinations thereof, wherein the Bacillus strain improves milk production and / or milk quality in the animal.

[0116] In another exemplary embodiment, a method of inhibiting a disease or disorder in a milk producing animal is provided, wherein the disease or disorder is selected from the group consisting of milk fever, ketosis, retained placenta, metritis, pneumonia, and ectopic abomasum. The method comprises the step of administering to the animal a feed composition or drinking water comprising an effective amount of an additive, wherein the additive comprises 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 the identifying characteristics of Bacillus strain 86 (NRRL No. B-50944), a strain having all the identifying characteristics of Bacillus strain 300 (NRRL No. B-50943), a strain having all the identifying characteristics of Bacillus strain 101 (NRRL No. B-67218), a strain having all the identifying characteristics of Bacillus strain 235 (NRRL No. B-67219), (NRRL No. B-67219), a strain having all the identifying characteristics of Bacillus strain 177 (NRRL No. B-67275), a strain having all the identifying characteristics of Bacillus strain 102 (NRRL No. B-67276), and combinations thereof, wherein the Bacillus strain inhibits a disease or disorder in a milk producing animal, wherein the disease or disorder is selected from the group consisting of milk fever, ketosis, retained placenta, metritis, pneumonia, and ectopic abomasum.

[0117] The following clauses and combinations thereof provide various additional exemplary aspects of the invention described herein. The various embodiments described in any other section of this patent application (including the section entitled "Background and Summary of the Invention" and the Examples) are applicable to any of the following embodiments of the invention described in the following numbered clauses.

[0118] 1. A method for improving the reproductive performance of an animal, the method comprising the step of administering to the animal a feed composition or drinking water comprising an effective amount of an additive, the 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 the identifying characteristics of Bacillus strain 86 (NRRL No. B-50944), a strain having all the identifying characteristics of Bacillus strain 300 (NRRL No. B-50943), a strain having all the identifying characteristics of Bacillus strain 101 (NRRL No. B-67218), a strain having all the identifying characteristics of Bacillus strain 235 (NRRL No. B-67219), a strain having all the identifying characteristics of Bacillus strain 177 (NRRL No. B-67275), a strain having all the identifying characteristics of Bacillus strain 102 (NRRL No. B-67276), and combinations thereof, wherein the Bacillus strain improves the reproductive performance of the animal.

[0119] 2. A method of reducing mortality losses and / or reducing fasting events in animals, the method comprising the step of administering to the animal a feed composition or drinking water comprising an effective amount of an additive, the 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 the identifying characteristics of Bacillus strain 86 (NRRL No. B-50944), a strain having all the identifying characteristics of Bacillus strain 300 (NRRL No. B-50943), a strain having all the identifying characteristics of Bacillus strain 101 (NRRL No. B-67218), a strain having all the identifying characteristics of Bacillus strain 235 (NRRL No. B-67219), a strain having all the identifying characteristics of Bacillus strain 177 (NRRL No. B-67275), a strain having all the identifying characteristics of Bacillus strain 102 (NRRL No. B-67276), and combinations thereof, wherein the Bacillus strain reduces mortality losses in animals and / or reduces fasting episodes in animals.

[0120] 3. The method of clause 1 or 2, wherein the animal is selected from poultry species, porcine species, bovine species, ovine species, equine species, and companion animals.

[0121] 4. The method of any one of clauses 1 to 3, wherein the animal is a porcine species, and the porcine species is selected from the group consisting of finishing pigs, nursery pigs, sows and breeder pigs.

[0122] 5. The method of clause 1 or 2, wherein the animal is a milk producing animal.

[0123] 6. The method of clause 5, wherein the milk-producing animal is selected from the group consisting of a cow, a goat, a sheep or a buffalo.

[0124] 7. The method of any one of clauses 1 to 6, wherein the Bacillus strain produces an enzyme selected from the group consisting of α-galactosidase, protease, lipase, amylase, xylanase, cellulase, and combinations thereof.

[0125] 8. The method of any one of clauses 1 to 7, further comprising the step of administering to the animal another bacterial strain selected from another Bacillus strain, a lactic acid bacteria strain, and combinations thereof.

[0126] 9. The method of any of clauses 1-8, 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.

[0127] 10. The method of any one of clauses 1 to 8, wherein the strain applied is Bacillus strain 101 (NRRL No. B-67218) or a strain having all the identifying characteristics of Bacillus strain 101 (NRRL No. B-67218).

[0128] 11. The method of any one of clauses 1 to 8, wherein the strain applied is Bacillus strain 235 (NRRL No. B-67219) or a strain having all the identifying characteristics of Bacillus strain 235 (NRRL No. B-67219).

[0129] 12. The method of any one of clauses 1 to 8, wherein the strain applied is Bacillus strain 86 (NRRL No. B-50944) or a strain having all the identifying characteristics of Bacillus strain 86 (NRRL No. B-50944).

[0130] 13. The method of any one of clauses 1 to 8, wherein the strain applied is Bacillus strain 300 (NRRL No. B-50943) or a strain having all the identifying characteristics of Bacillus strain 300 (NRRL No. B-50943).

[0131] 14. The method of any one of clauses 1 to 8, wherein the strain applied is Bacillus strain 177 (NRRL No. B-67275) or a strain having all the identifying characteristics of Bacillus strain 177 (NRRL No. B-67275).

[0132] 15. The method of any one of clauses 1 to 8, wherein the strain applied is Bacillus strain 102 (NRRL No. B-67276) or a strain having all the identifying characteristics of Bacillus strain 102 (NRRL No. B-67276).

[0133] 16. The method of any one of clauses 1 to 8, wherein the strain applied is Bacillus strain 101 (NRRL No. B-67218).

[0134] 17. The method of any one of clauses 1 to 8, wherein the strain applied is Bacillus strain 235 (NRRL No. B-67219).

[0135] 18. The method of any one of clauses 1 to 8, wherein the strain applied is Bacillus strain 86 (NRRL No. B-50944).

[0136] 19. The method of any one of clauses 1 to 8, wherein the strain applied is Bacillus strain 300 (NRRL No. B-50943).

[0137] 20. The method of any one of clauses 1 to 8, wherein the strain applied is Bacillus strain 177 (NRRL No. B-67275).

[0138] 21. The method of any one of clauses 1 to 8, wherein the strain applied is Bacillus strain 102 (NRRL No. B-67276).

[0139] 22. The method of any one of clauses 1 to 21, wherein the 3 CFU / lb of feed composition to about 5.0 x 10 12 Dosage of CFU / lb of feed composition Each Bacillus strain was administered in the feed composition.

[0140] 23. The method of any one of clauses 1 to 21, wherein the 7CFU / lb of feed composition to about 5.0 x 10 7 Dosage of CFU / lb of feed composition Each Bacillus strain was administered in the feed composition.

[0141] 24. The method of any one of clauses 1 to 21, wherein the 6 Dosage of CFU / lb of feed composition Each Bacillus strain was administered in the feed composition.

[0142] 25. The method of any one of clauses 1 to 24 further comprising the step of administering an antibiotic to the animal.

[0143] 26. The method of clause 25, wherein the antibiotic is selected from erythromycin, levofloxacin, trimethoprim / sulfamethoxazole, trimethoprim, daptomycin, rifampicin, tylosin, tilmicosin, vancomycin, monensin, lasalocid, and lelomycin propionate.

[0144] 27. The method of any one of clauses 1 to 26, further comprising the step of administering to the animal an enzyme selected from the group consisting of β-galactosidase, protease, lipase, amylase, hemicellulase, arabinoxylanase, xylanase, cellulase, NSPase, phytase, and combinations thereof.

[0145] 28. The method of clause 27, wherein the enzyme is NSPase or phytase.

[0146] 29. The method of clause 1 or 2, wherein the animal is a cow and the Bacillus strain is administered during lactation.

[0147] 30. The method of clause 1 or 2, wherein the animal is a cow and the Bacillus strain is administered during gestation.

[0148] 31. The method of any one of clauses 1 to 30, wherein the feed composition is administered to the animal daily.

[0149] 32. The method of any one of clauses 1 or 3-31, wherein the improvement in reproductive performance is selected from a reduction in the number of conception services, an increase in estrus detection, an increase in conception rate, a reduction in miscarriage and an increase in pregnancy rate.

[0150] 33. A method of improving milk production and / or milk quality in a dairy animal, the method comprising the step of administering to the animal a feed composition or drinking water comprising an effective amount of an additive, the 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 the identifying characteristics of Bacillus strain 86 (NRRL No. B-50944), a strain having all the identifying characteristics of Bacillus strain 300 (NRRL No. B-50943), a strain having Bacillus strain 101 (NRRL No. B-67218), (NRRL No. B-67218), a strain having all the identifying characteristics of Bacillus strain 235 (NRRL No. B-67219), a strain having all the identifying characteristics of Bacillus strain 177 (NRRL No. B-67275), a strain having all the identifying characteristics of Bacillus strain 102 (NRRL No. B-67276), and combinations thereof, wherein the Bacillus strain improves milk production and / or milk quality in the animal.

[0151] 34. A method of inhibiting a disease or disorder in a milk-producing animal, wherein the disease or disorder is selected from the group consisting of milk fever, ketosis, retained placenta, metritis, pneumonia, and ectopic abomasum, 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 the identifying characteristics of Bacillus strain 86 (NRRL No. B-50944), a strain having all the identifying characteristics of Bacillus strain 300 (NRRL No. B-50943), a strain having Bacillus strain 101 (NRRL No. B-67218), (NRRL No. B-67218), a strain having all the identifying characteristics of Bacillus strain 235 (NRRL No. B-67219), a strain having all the identifying characteristics of Bacillus strain 177 (NRRL No. B-67275), a strain having all the identifying characteristics of Bacillus strain 102 (NRRL No. B-67276), and combinations thereof, wherein the Bacillus strain inhibits a disease or disorder in the milk producing animal, wherein the disease or disorder is selected from the group consisting of milk fever, ketosis, retained placenta, metritis, pneumonia, and ectopic abomasum.

[0152] 35. The method of clause 33 or 34, wherein the milk producing animal is selected from a cow, goat, sheep or buffalo.

[0153] 36. The method of any one of clauses 33-35, wherein the Bacillus strain produces an enzyme selected from the group consisting of α-galactosidase, protease, lipase, amylase, xylanase, cellulase, and combinations thereof.

[0154] 37. The method of any one of clauses 33 to 36 further comprising the step of administering to the animal another bacterial strain selected from another Bacillus strain, a lactic acid bacteria strain, and combinations thereof.

[0155] 38. The method of any of clauses 33-37, 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.

[0156] 39. The method of any one of clauses 33 to 37, wherein the strain applied is Bacillus strain 101 (NRRL No. B-67218) or a strain having all the identifying characteristics of Bacillus strain 101 (NRRL No. B-67218).

[0157] 40. The method of any one of clauses 33 to 37, wherein the strain administered is Bacillus strain 235 (NRRL No. B-67219) or a strain having all of the identifying characteristics of Bacillus strain 235 (NRRL No. B-67219).

[0158] 41. The method of any one of clauses 33 to 37, wherein the strain administered is Bacillus strain 86 (NRRL No. B-50944) or a strain having all of the identifying characteristics of Bacillus strain 86 (NRRL No. B-50944).

[0159] 42. The method of any one of clauses 33 to 37, wherein the strain administered is Bacillus strain 300 (NRRL No. B-50943) or a strain having all of the identifying characteristics of Bacillus strain 300 (NRRL No. B-50943).

[0160] 43. The method of any one of clauses 33 to 37, wherein the strain administered is Bacillus strain 177 (NRRL No. B-67275) or a strain having all of the identifying characteristics of Bacillus strain 177 (NRRL No. B-67275).

[0161] 44. The method of any one of clauses 33 to 37, wherein the strain administered is Bacillus strain 102 (NRRL No. B-67276) or a strain having all of the identifying characteristics of Bacillus strain 102 (NRRL No. B-67276).

[0162] 45. The method of any one of clauses 33 to 37, wherein the strain applied is Bacillus strain 101 (NRRL No. B-67218).

[0163] 46. ​​The method of any one of clauses 33-37, wherein the strain applied is Bacillus strain 235 (NRRL No. B-67219).

[0164] 47. The method of any one of clauses 33-37, wherein the strain applied is Bacillus strain 86 (NRRL No. B-50944).

[0165] 48. The method of any one of clauses 33 to 37, wherein the strain applied is Bacillus strain 300 (NRRL No. B-50943).

[0166] 49. The method of any one of clauses 33-37, wherein the strain applied is Bacillus strain 177 (NRRL No. B-67275).

[0167] 50. The method of any one of clauses 33 to 37, wherein the strain applied is Bacillus strain 102 (NRRL No. B-67276).

[0168] 51. The method of any one of clauses 33 to 50, wherein the 3 CFU / lb of feed composition to approximately 5.0 x 10 12 Dosage of CFU / lb of feed composition Each Bacillus strain was administered in the feed composition.

[0169] 52. The method of any one of clauses 33 to 50, wherein the 7 CFU / lb of feed composition to approximately 5.0 x 10 7 Dosage of CFU / lb of feed composition Each Bacillus strain was administered in the feed composition.

[0170] 53. The method of any one of clauses 33-50, wherein the 6 Dosage of CFU / lb of feed composition Each Bacillus strain was administered in the feed composition.

[0171] 54. The method of any one of clauses 33 to 53 further comprising the step of administering an antibiotic to the animal.

[0172] 55. The method of clause 54, wherein the antibiotic is selected from erythromycin, levofloxacin, trimethoprim / sulfamethoxazole, trimethoprim, daptomycin, rifampin, tylosin, tilmicosin, vancomycin, monensin, lasalocid, and lelomycin propionate.

[0173] 56. The method of any one of clauses 33 to 55, further comprising the step of administering to the animal an enzyme selected from the group consisting of β-galactosidase, protease, lipase, amylase, hemicellulase, arabinoxylanase, xylanase, cellulase, NSPase, phytase, and combinations thereof.

[0174] 57. The method of clause 56, wherein the enzyme is NSPase or phytase.

[0175] 58. The method of any one of clauses 33 to 57, wherein the Bacillus strain is administered during lactation.

[0176] 59. The method of any one of clauses 33-57, wherein the Bacillus strain is administered during pregnancy.

[0177] 60. The method of any one of clauses 33-59, wherein the feed composition is administered to the animal daily.

[0178] 61. The method of any one of clauses 33 or 35-60 wherein the improvement in milk yield and / or milk quality is selected from: increasing milk yield, increasing milk fat percentage, increasing milk protein, increasing total fat corrected milk, increasing energy corrected milk yield and reducing somatic cell count.

[0179] 62. The method of any one of clauses 1, 2 or 5-61 wherein the animal is a cow.

[0180] 63. A commercial package for animal use comprising all isolated strains including 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] 64. A feed additive for animal feed comprising all isolated strains, wherein the isolated strains include 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] 65. An additive for animal drinking water comprising all isolated strains, wherein the isolated strains include 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).

[0183] 66. An animal feed composition comprising all isolated strains including 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).

[0184] 67. A commercial package, a feed additive, an additive for drinking water for an animal or an animal feed composition according to any one of clauses 63 to 66 for improving the reproductive performance of said animal.

[0185] 68. A commercial package, a feed additive, an additive for drinking water for animals or an animal feed composition as claimed in any one of clauses 63 to 66 for use in reducing mortality losses and / or fasting events in animals.

[0186] 69. A commercial package, a feed additive, an additive for drinking water for animals or an animal feed composition according to any of clauses 63 to 66 for improving milk production and / or milk quality in an animal.

[0187] 70. A commercial package, a feed additive, an additive for animal drinking water or an animal feed composition as claimed in any one of clauses 63 to 66 for use in inhibiting a disease or disorder in an animal selected from the group consisting of: lactogenic fever, ketosis, retained placenta, metritis, pneumonia and ectopic abomasum.

[0188] 71. A feed additive or an additive for drinking water for animals as defined in item 64 or 65 in the form of a concentrate.

[0189] 72. A feed additive or an additive for drinking water for animals as defined in item 64 or 65 in the form of a super concentrate.

[0190] 73. A feed additive, a feed composition or an additive for drinking water for animals according to any one of clauses 64 to 72 in dry form.

[0191] 74. The feed composition of clause 73 in the form of pellets.

[0192] 75. A commercial package, a feed additive or an additive for drinking water for animals according to any one of clauses 63-65 or 67-74, wherein the strain is in a form selected from the group consisting of a powder, a liquid and a pelleted form.

[0193] 76. A commercial package, a feed additive, an additive for drinking water for animals or a feed composition according to any one of clauses 63 to 75, further comprising a carrier of said Bacillus strain.

[0194] 77. The commercial package, feed additive, additive for animal drinking water or feed composition of clause 76, wherein the carrier is selected from the group consisting of bran, rice hulls, salt, dextrin and combinations thereof.

[0195] 78. A commercial package, a feed additive, an additive for drinking water for animals or a feed composition according to any of clauses 63 to 77 in a bag.

[0196] 79. The commercial bag, feed additive, additive for drinking water for animals or feed composition of clause 78, wherein the bag is a plastic bag.

[0197] 80. The commercial package, feed additive, additive for drinking water for animals or feed composition of any one of clauses 63 to 79, further comprising instructions for use of the one or more Bacillus strains.

[0198] 81. A commercial package, feed additive, feed composition, or additive for drinking water for animals of any of clauses 63 to 80 in a 20-pound bag.

[0199] 82. A commercial package, feed additive, feed composition, or additive for drinking water for animals of any of clauses 63 to 80 in a 50-pound bag.

[0200] 83. A feed additive or an additive for drinking water for animals according to any of clauses 64, 65, 67-73 or 75-82 in powder form.

[0201] 84. A feed additive or an additive for drinking water for animals according to any of clauses 64, 65, 67-72, 75 or 78-80 in liquid form.

[0202] 85. A commercial package, a feed additive, an additive for drinking water for animals or a feed composition as claimed in any one of clauses 63 to 84 in a container for commercial use.

[0203] 86. The commercial package, feed additive, additive for drinking water for animals or feed composition of clause 85, wherein the container comprises plastic.

[0204] 87. The commercial bag, feed additive, additive for drinking water for animals or feed composition of clause 85, wherein the container comprises paper.

[0205] 88. A commercial package, a feed additive, an additive for drinking water for animals or a feed composition according to any one of clauses 63 to 87, wherein the strain further comprises a binder.

[0206] 89. The commercial package, feed additive, additive for animal drinking water or feed composition of clause 88, wherein the binder is selected from clay, yeast cell wall components, aluminum silicate and dextran or a combination thereof.

[0207] In different embodiments, the animal to which the feed additive, feed composition or drinking water as described herein is applied can be selected from poultry species, pig species, cattle species, sheep species, horse species and companion animals. In the embodiment where the animal is a companion animal, the companion animal can be, for example, canine species or feline species. In the embodiment where the animal is a pig species, the pig species can be selected from growing and completing pigs, nursery pigs, sows and breeding pigs. In different exemplary embodiments, the animal can be selected from chicken (for example, broiler or laying hen), pig, horse, pony, cow, buffalo, turkey, goat, sheep, quail, pheasant, ostrich, duck and their combination. In an exemplary embodiment, the animal can be a cow.

[0208] In any embodiment described herein except those that need to be combined to apply all bacterial strains, the bacterial strain can be applied individually or in any combination.In one embodiment, the bacillus strain of effective amount can be applied to animals.“Effective amount” refers to the amount of the bacillus strain (e.g., bacterial strain 86, 300, 101 or 235, 177 or 102, or bacterial strains with all their identification characteristics) that can improve the reproductive performance of animals, reduce the death loss and / or fasting events of animals, improve the milk production and / or milk quality of dairy animals and / or suppress the disease or obstacle of dairy animals as described below, the disease or obstacle is selected from lactogenic fever, ketosis, placenta retention, metritis, pneumonia and ectopic abomasum.In one aspect, the improvement of reproductive performance is selected from the reduction of pregnancy service number of times, the increase of estrus detection rate, the increase of conception rate, the reduction of abortion and the increase of pregnancy rate (e.g., 21-day pregnancy rate). In another exemplary aspect, the improvement in milk yield and / or milk quality is selected from the group consisting of increasing milk yield, increasing milk fat percentage, increasing milk protein, increasing total fat-corrected milk, increasing energy-corrected milk yield, and reducing somatic cell counts (e.g., reducing the log mean of somatic cell counts).

[0209] As used herein, "inhibiting a disease or disorder" may refer to reducing the number of animals with a disease or disorder, preventing a disease or disorder, alleviating the symptoms of a disease or disorder in an animal, or a combination thereof, wherein the disease or disorder is selected from lactogenic fever, ketosis, retained placenta, metritis, pneumonia, and ectopic abomasum. "Reducing animal death losses" as used herein may refer to reducing the number of animal losses caused by death, or preventing animal losses caused by death. "Reducing animal fasting events" as used herein may refer to reducing the number of animal fasting events, or preventing any fasting events of animals, or, for example, increasing daily rumination (e.g., the time of daily rumination). The terms "inhibit", "reduce", "increase", "improve" or "reduce" as used herein, for example, are all relative to the condition of an animal (e.g., a dairy animal) before administering one or more strains to the animal (named "before Bacillus" in the examples), or, for example, relative to an animal not fed with a Bacillus strain. As described herein, "improving milk quality" as used herein can refer to increasing milk fat percentage (or, for example, pounds of milk fat), increasing milk protein (e.g., in percentage or pounds), increasing total fat corrected milk, increasing energy corrected milk production and / or reducing somatic cell count. "Energy corrected milk" (ECM) as used herein is a calculation of the amount of energy in milk based on milk volume, fat and protein. Energy corrected milk determines the amount of milk produced, which is adjusted to 3.5% fat and 3.2% protein. Energy corrected milk is a more useful measurement than simple milk production volume to track herd productivity because it puts all animals on an equal footing for comparison purposes over time.

[0210] In the embodiments described herein where the compositions of the 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 the animal 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 an exemplary embodiment, 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 an animal.

[0211] In another exemplary embodiment, 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 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, mixed with an animal feed blend, including any animal feed blend known in the art or any animal feed blend described herein. As used herein, "feed composition" or "animal feed composition" refers to 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, mixed with an animal feed blend, and optionally with any other components that may be used in a feed composition, including other bacterial strains, such as other Bacillus strains or Lactobacillus ( Lactobacillus ) strains.

[0212] According to the method and composition described in this patent application, any animal feed admixture can be used, including those known in the art and those described herein, such as rapeseed meal, cottonseed meal, soybean meal, corn meal, barley, wheat, silage and silage hay. In different embodiments, a full mixed diet based on corn silage and alfalfa hay can be used, or a corn silage mixed diet can be used. In different embodiments, 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 a strain with all their identification characteristics can be supplemented to the animal feed admixture, but other ingredients can be optionally added to the animal feed admixture, including other different bacterial strains, such as other bacillus strains or lactobacillus strains.

[0213] In various exemplary embodiments, the optional ingredients of the animal feed blend include sugars and complex sugars such as water-soluble and water-insoluble monosaccharides, disaccharides and polysaccharides. Other optional ingredients include dried distiller's grains, grain solubles, fats (e.g., crude fat), phosphorus, sodium bicarbonate, limestone, salt, phytate, calcium, sodium, sulfur, magnesium, potassium, copper, iron, manganese, zinc, ash, fish oil, oil derived from fish meal, raw seeds (e.g., linseed), antioxidants and starch. In another embodiment, minerals can be added in the form of a mineral premix.

[0214] The optional amino acid components that can be added to the animal feed admixture are arginine, histidine, isoleucine, leucine, lysine, cysteine, methionine, phenylalanine, threonine, tryptophan, valine, tyrosine ethyl ester hydrochloride, alanine, aspartic acid, sodium glutamate, glycine, proline, serine, cysteine ​​ethyl ester hydrochloride and the like, and salts thereof. The vitamins that can be optionally added are thiamine hydrochloride, riboflavin, pyridoxine hydrochloride, nicotinic acid, nicotinamide, inositol, choline chloride, calcium pantothenate, biotin, folic acid, ascorbic acid, and vitamins A, B, K, D, E, etc. In another embodiment, vitamins can be added in the form of a vitamin premix. In yet another embodiment, protein components can be added to the animal feed admixture and include protein obtained from meat meal, bone meal or fish meal, liquid or powdered eggs, fish solubles, crude protein, etc.

[0215] In another exemplary aspect, any pharmaceutical ingredient known in the art can be added to an animal feed admixture or to an additive for animal drinking water, such as an antibiotic. In different embodiments, the antibiotic is selected from erythromycin, levofloxacin, trimethoprim / sulfamethoxazole, trimethoprim, daptomycin, rifampicin, tylosin, tilmicosin, vancomycin, monensin, lasalocid and lelomycin propionate and combinations thereof. In another embodiment, an animal feed admixture, a feed composition, a feed additive or an additive for animal drinking water may contain an antibiotic or may not contain an antibiotic.

[0216] In another exemplary embodiment, one or more enzymes may be added to the animal feed blend. In various embodiments, the enzymes that may be added include β-galactosidase, phytase, protease, lipase, amylase, hemicellulase, arabinoxylanase, xylanase, cellulase, NSPase, combinations thereof, and any other enzymes that improve the effectiveness of the feed composition for the uses described herein. In yet another embodiment, yeast, fungi (e.g., Aspergillus or Trichoderma ( Trichoderma )) or micronutrients are added to animal feed. Any of the above ingredients suitable for addition to an additive for animal drinking water can be added as a component of an additive for animal drinking water as described herein.

[0217] In various exemplary embodiments, a 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 strain added in addition to these Bacillus strains can be administered in the feed composition, each strain at about 1.0 x 10 3 CFU / lb of feed composition to approximately 5.0 x 10 12 The dose of CFU / lb of feed composition was about 0.5 x 10 7 CFU / lb of feed composition to approximately 5.0 x 10 7 CFU / lb of feed composition, or at a dose greater than about 5.0 x 10 6 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 a feed composition at a dose greater than about 1.0 x 10 3 CFU / lb of feed composition at a dose greater than about 1.1 x 10 3CFU / lb of feed composition at a dose greater than about 1.25 x 10 3 CFU / lb of feed composition at a dose greater than about 1.5 x 10 3 CFU / lb of feed composition at a dose greater than about 1.75 x 10 3 CFU / lb of feed composition at a dose greater than about 1.0 x 10 4 CFU / lb of feed composition at a dose greater than about 2.0 x 10 4 CFU / lb of feed composition at a dose greater than about 3.0 x 10 4 CFU / lb of feed composition at a dose greater than about 4.0 x 10 4 CFU / lb of feed composition at a dose greater than about 5.0 x 10 4 CFU / lb of feed composition at a dose greater than about 6.0 x 10 4 CFU / lb of feed composition at a dose greater than about 7.0 x 10 4 CFU / lb of feed composition at a dose greater than about 8.0 x 10 4 CFU / lb of feed composition at a dose greater than about 1.0 x 10 5 CFU / lb of feed composition at a dose greater than about 1.0 x 10 6 CFU / lb of feed composition at a dose greater than about 2.0 x 10 6 CFU / lb of feed composition at a dose greater than about 3.0 x 10 6 CFU / lb of feed composition at a dose greater than about 4.0 x 10 6 CFU / lb of feed composition at a dose greater than about 5.0 x 10 6 CFU / lb of feed composition at a dose greater than about 6.0 x 10 6 CFU / lb of feed composition at a dose greater than about 7.0 x 10 6 CFU / lb of feed composition at a dose greater than about 8.0 x 10 6 CFU / lb of feed composition at a dose greater than about 9.0 x 10 6 CFU / lb of feed composition at a dose greater than about 1.0 x 10 7 CFU / lb of feed composition at a dose greater than about 2.0 x 10 7 CFU / lb of feed composition at a dose greater than about 3.0 x 10 7 CFU / lb of feed composition at a dose greater than about 4.0 x 10 7CFU / lb of feed composition at a dose greater than about 5.0 x 10 7 CFU / lb of feed composition at a dose greater than about 1.0 x 10 8 CFU / lb of feed composition at a dose greater than about 1.0 x 10 9 CFU / lb of feed composition at a dose greater than about 1.0 x 10 10 CFU / lb of feed composition at a dose greater than about 1.0 x 10 11 CFU / lb of feed composition, or a dose thereof greater than about 1.0 x 10 12 CFU / lb of feed composition. In all of the embodiments described in this paragraph, the animal can be fed or the animal 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.

[0218] In other embodiments, the Bacillus strains (e.g., Bacillus strains 86, 300, 101, 235, 177, and 102 and / or strains having all of their identifying characteristics) can be administered in a feed composition by adding about 15 grams of a feed additive to a complete feed composition, wherein the 15 grams of the additive contains all of the strains to provide about 1.0 x 10 of the total amount of feed to be fed to the animal per day in the 15 grams of the additive. 9 CFU, about 2.0 x 10 9 CFU, about 3.0 x 10 9 CFU, about 4.0 x10 9 CFU, about 5.0 x 10 9 CFU, about 6.0 x 10 9 CFU, about 7.0 x 10 9 CFU, about 8.0 x 10 9 CFU, about 9.0 x10 9 CFU or approximately 1.0 x 10 10 Total combined strains of CFU. In all embodiments described in this paragraph, the animal can be fed or the animal 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 15 grams of feed additive can be added to any of these amounts of the feed composition.

[0219] In another embodiment, by mixing about 15 grams of 9 CFU of the combined strains (e.g., a total of approximately 7.35 x 10 9CFU) is added to a complete feed composition, for example, 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 to be fed or consumed per animal per day. The Bacillus strain (e.g., Bacillus strains 86, 300, 101, 235, 177, and 102 and / or strains having all of their identifying characteristics) can be administered in the feed composition.

[0220] In any of the dosage embodiments of the above-described Bacillus strains 86, 300, 101, 235, 177 and 102 and / or strains having all of their identifying characteristics, the Bacillus strains (Bacillus strains 86, 300, 101, 235, 177 and 102 and / or strains having all of their identifying characteristics) may be included in a feed additive for addition to a feed composition in an exemplary ratio selected from any of the following ratios, wherein the ratios refer, in that order, to Bacillus strains 86, 300, 101, 235, 177 and 102 and / or strains having all of their identifying characteristics. 1:1:1:1:1:1 1:0.8:1:1:1:1 1:1:0.8:1:1:1 1:1:1:0.8:1:1 1:1:1:1:0.8:1 1:1:1:1:1:0.8 1:0.5:1:1:1:1 1:1:0.5:1:1:1 1:1:1:0.5:1:1 1:1:1:1:0.5:1 1:1:1:1:1:0.5 1:0.2:1:1:1:1 1:1:0.2:1:1:1 1:1:1:0.2:1:1 1:1:1:1:0.2:1 1:1:1:1:1:0.2 0.8:0.5:1:1:1:1 1:0.8:0.5:1:1:1 1:1:0.8:0.5:1:1 1:1:1:0.8:0.5:1 1:1:1:1:0.8:0.5 0.2:0.5:1:1:1:1 1:0.2:0.5:1:1:1 1:1:0.2:0.5:1:1 1:1:1:0.2:0.5:1 1:1:1:1:0.2:0.5 0.8:0.2:1:1:1:1 1:0.8:0.2:1:1:1 1:1:0.8:0.2:1:1 1:1:1:0.8:0.2:1 1:1:1:1:0.8:0.2 0.8:0.5:0.2:1:1:1 1:0.8:0.5:0.2:1:1 1:1:0.8:0.5:0.2:1 1:1:1:0.8:0.5:0.2 0.2:1:1:1:0.8:0.5 0.8:0.2:0.5:1:1:1 1:0.8:0.2:0.5:1:1 1:1:0.8:0.2:0.5:1 1:1:1:0.8:0.2:0.5 0.5:1:1:1:0.8:0.2 0.2:0.8.:0.5:1:1:1 1:0.2:0.8:0.5:1:1 1:1:0.2:0.8:0.5:1 1:1:1:0.2:0.8:0.5

[0266] 0.2:1:1:1:0.8:0.5。

[0267] In various embodiments, the Bacillus strains for use according to the methods and compositions described herein (e.g., Bacillus strain 86, and / or 300, and / or 101, and / or 235, and / or 177, and / or 102) can be selected from 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 Bacillus strain 101 (NRRL No. B-67218), Bacillus strain MDG 101 and Bacillus strain MDG 235 were deposited on January 4, 2016 at the Agricultural Research Service Culture Collection (NRRL), National Centerfor Agricultural Utilization Research, Agricultural Research Service, USDA, 1815 North University Street, Peoria, Illinois 61604-3999, 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 strains 86 and 300, respectively, as referred to in this application. The NRRL strain designations MDG 101, MDG 235, MGL177, and MGL102 are equivalent to Bacillus strains 101, 235, 177, and 102, respectively, as referred to in this application. The deposit was 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 may 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.

[0268] Bacillus strains 102 and 177 are Bacillus pumilus ( Bacillus pumilus ) strains, and Bacillus strain 77 is Bacillus licheniformis ( Bacillus licheniformis ) strains, and all other strains described in this application are Bacillus subtilis ( Bacillus subtilus ) strain.

[0269] 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.

[0270] In another embodiment, one or more bacillus strains described in the above 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 with all their identification characteristics) can be administered to animals together with another different bacterial strain, wherein the another different bacterial strain is selected from another bacillus strain, a lactic acid bacteria strain, and a combination thereof. In yet another embodiment, one or more bacillus strains described in the above 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 with all their identification characteristics) can be administered to animals together with any other different bacterial strains effective for the purposes described herein.

[0271] As used herein, a “strain having all 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 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 mutant strain having a similarity to Bacillus strain 86, Bacillus strain 300, Bacillus strain 101, Bacillus strain 235, Bacillus strain 177, or Bacillus strain 102 based on DNA analysis). 02, enzyme activities corresponding to Bacillus strain 86, Bacillus strain 300, Bacillus strain 101, Bacillus strain 235, Bacillus strain 177 or Bacillus strain 102, antimicrobial activities corresponding to Bacillus strain 86, Bacillus strain 300, Bacillus strain 101, Bacillus strain 235, Bacillus strain 177 or Bacillus strain 102, antibiotic sensitivity and tolerance profiles corresponding to Bacillus strain 86, Bacillus strain 300, Bacillus strain 101, Bacillus strain 235, Bacillus strain 177 or Bacillus strain 102). In alternative embodiments, the mutations may be natural mutations, or genetically engineered mutations. In another embodiment, a "strain having all 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, for example, a strain 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 bacteria, such as another Bacillus strain, so 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 that corresponds based on DNA analysis to the DNA fingerprint of Bacillus Strain 86, Bacillus Strain 300, Bacillus Strain 101, Bacillus Strain 235, Bacillus Strain 177, or Bacillus Strain 102).

[0272] Feed compositions 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 with all of their identifying characteristics can be administered to animals for any time period or combination thereof effective for the purposes described herein. For example, in one embodiment, the feed composition or drinking water can be provided to the animal every day. In an alternative embodiment, the feed composition or drinking water can be administered to animals during lactation and / or during pregnancy. The administration time periods of the above-mentioned feed compositions or drinking water are non-limiting examples, and it should be understood that any time period or administration plan or combination thereof determined to be effective for the purposes described herein can be used.

[0273] In another embodiment of the invention, compositions are provided 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 one embodiment, commercial packages are provided for animal use, comprising all of the isolated strains, including 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).

[0274] In another embodiment, a feed additive for animal feed is provided, comprising all isolated strains including 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).

[0275] In yet another embodiment, an additive for animal drinking water is provided, comprising all of the isolated strains, including 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).

[0276] In yet another embodiment, an animal feed composition is provided that comprises all of the isolated strains including 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).

[0277] In one embodiment, the feed additive for addition to the animal feed blend to produce a complete feed composition can be mixed with the animal feed blend, for example, with an automated micronutrient delivery system, or, for example, by manual 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 an animal in the form of a complete feed composition. The mixing can also be accomplished by any other suitable method known in the art for combining a direct-fed microorganism with an animal feed blend to obtain a homogenous mixture. In various embodiments, the mixing can be performed for any suitable period of time (e.g., from about 1 to about 4 minutes). In embodiments where 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 the form of an additive for drinking water for animals, 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 be in the form of, for example, a powder, liquid, or pellets, and can be mixed with 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 an animal in the animal's drinking water. 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 orally (i.e., by oral insertion) directly administered to an animal in the form of a powder, liquid, capsule, top-dressing, gel, or pellet.

[0278] 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 improving the reproductive performance of the animal, reducing death losses and / or fasting events in the animal, improving milk production and / or milk quality in the animal, and / or inhibiting a disease or disorder in the animal selected from milk fever, ketosis, retained placenta, metritis, pneumonia, and ectopic abomasum.

[0279] In one exemplary aspect, the feed additive, the additive for animal drinking water, or the feed composition can be in the form of a commercial package. In another exemplary embodiment, the feed additive or the additive for animal drinking water containing 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 a commercial package can be in the form of a commercial package containing, for example, about 1 x 10 8 About 7 x 10 10 CFU / g of a combined strain concentrate or super concentrate (e.g., about 8 x 10 10 About 5 x 10 12 In another embodiment, the feed additive, feed composition or additive for animal drinking water, or 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 dry form (e.g., powder), pellet form, liquid form, in topping form or in gel form, in capsule form, or in any other suitable form.

[0280] In yet another embodiment, the strain in commercial package form can be, for example, in dry form (eg, powder or freeze-dried form), in pellet form, or in liquid form.

[0281] In another exemplary embodiment, the commercial bag, feed additive, additive for animal drinking water or feed composition can further include bacillus strain 86, bacillus strain 300, bacillus strain 101, bacillus strain 235, bacillus strain 177 and bacillus strain 102, and / or a carrier of strains with all their identification characteristics. The carrier can be selected from bran, rice husk, salt, mineral oil, dextrin (e.g., maltodextrin), whey, sugar, limestone, dry starch, sodium aluminosilicate, vegetable oil and their combination. In another embodiment, the carrier can be any suitable carrier for direct feeding microorganisms known in the art. In another embodiment, the commercial bag, feed additive, additive for animal drinking water or feed composition can further include adhesives such as clay, yeast cell wall components, aluminum silicate, dextran or other known adhesives. In another embodiment, the commercial bag, feed additive, additive for animal drinking water or feed composition can further include inorganic / organic adhesives, essential oils and / or organic acids. Any binder or carrier components may be added exogenously (ie, not naturally present in combination with the Bacillus strain), and any composition described herein may be present in a dietary nutritional composition (eg, a probiotic composition).

[0282] In other embodiments again, commercial bag, feed additive, additive for animal drinking water or feed composition comprising bacillus strain 86, bacillus strain 300, bacillus strain 101, bacillus strain 235, bacillus strain 177 and bacillus strain 102 and / or the bacterial strain with all their identification characteristics are in the container for commercial use.In different embodiments, the container can be such as bag (for example, 20-pound bag, 50-pound bag, 2-ounce bag, 1-pound bag or 1-kilogram bag), sac, cylinder, bottle or box.In exemplary aspects, commercial bag, feed additive, additive for animal drinking water or feed composition comprising bacillus strain 86, bacillus strain 300, bacillus strain 101, bacillus strain 235, bacillus strain 177 and bacillus strain 102 and / or the bacterial strain with all their identification characteristics can comprise plastics, metal, foil, paper, fiber or cardboard (for example, plastic bucket, paper bag, foil bag, fiber cylinder etc.). The commercial package, feed additive, additive for drinking water for animals or feed composition may further comprise instructions for use of the one or more Bacillus strains.

[0283] In one aspect, any composition described herein can further comprise exogenously added (i.e., not naturally present with the Bacillus strain) nutritional components selected from vitamins, antibiotics, enzymes, water-soluble or water-insoluble monosaccharides, disaccharides or polysaccharides, fats, phosphorus, sodium bicarbonate, limestone, calcium, sodium, sulfur, magnesium, potassium, copper, iron, manganese, zinc, fish oil, raw seeds, antioxidants and starch. In one embodiment, the exogenously added nutritional component can be an enzyme, and the enzyme can be selected from β-galactosidase, protease, lipase, amylase, hemicellulase, arabinoxylanase, xylanase, cellulase, NSPase, phytase and combinations thereof.

[0284] The following examples are for illustrative purposes only. The examples are non-limiting and are not intended to limit the present invention in any way.

[0285] Example 1

[0286] Field trials

[0287] The purpose of the field trial was to determine the effects of supplementation with 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 cow health, milk production, and reproductive parameters on a 700-cow Upper Midwestern Holstein dairy farm with an above average incidence of digestive problems.

[0288] Experimental Design and Methods

[0289] The 3-month period before Bacillus supplementation served as a baseline, and production, reproduction, and health indicators of the entire herd were monitored by Dairy Comp 305 (ValleyAgricultural Software, Tulare, CA). The entire herd was then monitored for milk production, milk components, health, and reproductive performance for 10 months, during which cows received daily Bacillus supplementation, and weekly single cow milk production, monthly DHIA milk components, and monthly health events were measured. During the Bacillus supplementation period, cows received 15 grams of Bacillus product per day to provide 7.35 x 10 per head per day in a standard total mixed diet based on corn silage and alfalfa hay. 9Colony forming units (CFU) of viable Bacillus. Reproduction indices were evaluated over a 6-month period in consecutive years, during which time the pre-supplementation with Bacillus was compared with the post-supplementation to control for seasonal variations.

[0290] Statistical analysis

[0291] Production and composition data were analyzed by the MIXED procedure of SAS (version 9.2 SAS Institute, Cary NC), with days in milk (DIM) as a covariate, looking for fixed effects of treatment and treatment*month*year interactions. Health and reproductive indices were evaluated by the GLIMMIX procedure of SAS. Significance between means was defined at p ≤ 0.05, and trends were indicated when p ≤ 0.1.

[0292] result

[0293] Feeding 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) reduced the incidence of ectopic abomasum, ketosis, and metritis in postpartum cows.

[0294] Table 1. Effect of treatment on mean percentage of postpartum cows experiencing illness.

[0295]

[0296] Feeding 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) increased milk fat percentage and total energy corrected milk in the entire herd.

[0297] Table 2. Effects of treatments on mean percentages of milk components and daily energy-corrected milk production .

[0298]

[0299] Feeding 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) improved reproductive indicators in the herd as evidenced by fewer services per conception, higher estrus detection, conception rate, and 21-day pregnancy rate.

[0300] Table 3. Effects of treatments on reproductive efficiency indicators.

[0301]

[0302] Example 2

[0303] Field trials

[0304] The purpose of the field trial was to determine the effects of supplementation with 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 rectal microbial populations, cow health, and milk production in a 14,000-cow dairy in the Mountain-West region of the United States with a higher than average incidence of digestive problems and a history of jejunal hemorrhagic syndrome.

[0305] Experimental Design and Methods

[0306] Direct-fed Bacillus microorganisms were supplemented to dairy cows in Idaho for a period of 6 months. During the Bacillus supplementation period, cows received 15 grams of the Bacillus product per day to provide 7.35 x 10 per head per day in a standard total mixed diet based on corn silage and alfalfa-silage haylage. 9 Colony forming units (CFU) of live Bacillus. Before DFM supplementation, 40 randomly selected cows were sampled by rectal scraping to establish baseline amounts of gastrointestinal-related bacterial and fungal markers and virulence genes. The same host animals (n=37) were followed and the sampling was repeated 60 days after the implementation of Bacillus supplementation. Genomic DNA was isolated from fresh rectal swabs, quantified, and qPCR was performed for quantification of target genes using the primers and qPCR conditions mentioned in Table 4.

[0307] Table 4. Microorganisms and associated gene targets measured by quantitative PCR (qPCR).

[0308]

[0309] The period of the first 6 months of bacillus supplementation served as a baseline, and production and health indicators were monitored for the entire herd by Dairy Comp 305 (ValleyAgricultural Software, Tulare, CA). Then, for the next 6 months, the entire herd received daily bacillus supplementation, and milk production and health events were monitored using weekly single cow milk production and monthly health events. Milk fat and protein were evaluated monthly for a subset of cows (n = 1055) in the same time period.

[0310] Statistical analysis

[0311] Microbial gene abundance was log-quantitated using one-way ANOVA and Tukey pairwise comparisons in Minitab v.17 (State College, PA). 10 The significant effects of Bacillus treatment were transformed and analyzed. Significance between means was defined at p ≤ 0.05, and trends were indicated at p ≤ 0.1. Production and component data were analyzed by the MIXED procedure of SAS (version 9.2 SAS Institute, Cary NC), with days in milk (DIM) as a covariate, looking for fixed effects of treatment and treatment*month*year interactions. Health and reproductive indices were evaluated by the GLIMMIX procedure of SAS. Significance between means was defined at p ≤ 0.05, and trends were indicated at p ≤ 0.1.

[0312] result

[0313] Feeding 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) reduced gastrointestinal carriage of undesirable microbial populations. Specifically, longitudinal tracking of individual cows revealed that after 60 days of Bacillus supplementation, total enteropathogenic and enterohemorrhagic E. coli were reduced. (16S)、 Heat-labile enterotoxin gene ( LT ) and tight adhesion and elimination genes ( eae ) was significantly reduced in the rectum. 18S) genes were also reduced after supplementation (p<0.05), and the α and β toxin genes of Clostridium perfringens (respectively CPA and CPB ) decreased numerically during the Bacillus supplementation process ( Figure 1 Feeding 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) resulted in a reduced incidence of milk fever (hypocalcemia) in postpartum cows (Table 5) as well as a reduction in the number of cows fasting in the entire herd (Table 5).

[0314] Table 5. Effect of treatment on mean percentage of cows experiencing disease.

[0315]

[0316] Feeding 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) resulted in significantly increased milk fat and protein percentages (Table 6), which in turn improved energy-corrected milk yield (Table 6), compared to the first 6 months of Bacillus supplementation.

[0317] Table 6. Effects of treatments on mean percentages of milk components and daily energy-corrected milk production .

[0318]

[0319] Example 3

[0320] Meta-analysis of 5 field trials

[0321] The objective of this study was to determine the effects of supplementation of five U.S. dairy herds with 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 transition cow health, herd mortality losses, abortion rates, milk quality, milk composition, and milk production.

[0322] Experimental Design and Methods

[0323] Production, reproduction, and health indicators were monitored in five herds representing 20,000 cows using Dairy Comp 305 (ValleyAgricultural Software, Tulare, CA) or PCDART (DRMS, Raleigh, NC) software, using a 3-month period prior to Bacillus supplementation as a baseline. The herds were then monitored for 3 months, during which cows received daily Bacillus supplementation, and weekly individual cow milk production, monthly DHIA milk components, and monthly health events were measured. During the Bacillus supplementation period, cows received 15 grams of Bacillus product per day to provide 7.35 x 10 per head per day in a total mixed ration based on corn silage. 9 Colony forming units (CFU) of live Bacillus.

[0324] Statistical analysis

[0325] Milk production and composition data were analyzed with the MIXED procedure of SAS using days in milk and farm as covariates, fixed effects of treatment, and interactions of treatment*month*year. Health and reproductive indices were evaluated with the GLIMMIX procedure of SAS.

[0326] result

[0327] Feeding 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) resulted in significantly reduced incidences of retained placenta and ketosis, which are important health challenges in transition dairy cows (Table 7). Total herd mortality losses were also lower during the Bacillus supplementation period.

[0328] Table 7. Effects of treatments on mean percentage of disease events and total herd mortality losses during the transition period .

[0329]

[0330] Feeding 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) resulted in significantly increased milk fat and protein percentages (Table 8). During Bacillus consumption, energy-corrected milk production increased by 2.47 pounds / cow / day. In addition, somatic cell counts, indicators of bacterial infection of the mammary gland, and measures of milk quality also decreased during Bacillus supplementation (Table 8).

[0331] Table 8. Effects of treatments on mean milk somatic cell counts, percentages of milk components, and daily energy-corrected milk production.

[0332]

[0333] Feeding 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) improved reproductive indicators in the herd, as evidenced by fewer abortions (Table 9).

[0334] Table 9. Effect of treatment on mean percentage of miscarriage.

[0335]

[0336] Example 4

[0337] RAPD (random amplified polymorphic DNA) analysis of DFM strains

[0338] To obtain the genetic barcode or "fingerprint" of the Bacillus strains described in this article, RAPD (random amplified polymorphic DNA) analysis was performed on each strain. Each isolate was cultured overnight at 32°C in 10 ml of tryptic soy broth. Thereafter, DNA was extracted from these cultures using Qiagen's DNeasy Mini kit according to the protocol provided for Gram-positive bacteria. RAPD-PCR was performed on each DNA sample using the GE Healthcare Illustra Ready-To Go RAPD kit to amplify gene fragments of any length. The RAPD primers used for RAPD-PCR analysis included the following: Primer 1 (5'-GGTGCGGGAA-3); Primer 2 (5'-GTTTCGCTCC3'); and Primer 3 (5'-GTAGACCCGT-3). PCR products were separated by gel electrophoresis, and the banding patterns were detected and matched using BioNumerics software. As shown in Figure 2-6 As shown in , no common DNA fingerprint was observed between DFM strains. Figure 2-6 It was confirmed that the DFM strains described herein are different strains, each with a unique DNA fingerprint.

[0339] Example 5

[0340] Field trials

[0341] The purpose of this field trial was to determine the effects of supplementation with 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 reproductive parameters of cows on an 870-cow Central Texas Holstein dairy farm with average reproductive efficiency indicators.

[0342] Experimental Design and Methods

[0343] Reproductive parameters were monitored throughout the herd using the Dairy Comp 305 (Valley Agricultural Software, Tulare, CA). Cows received daily Bacillus supplementation at a rate of 7.5 g of Bacillus product to provide 3.68 x 10 per head per day on a standard total mixed ration based on corn silage, hay, and haylage. 9Colony forming units (CFU) of live Bacillus. Conception rate, service per conception, 21-day pregnancy rate and estrus detection rate were calculated and measured over a 6-month period (November to April) after the start of Bacillus supplementation. The same 6-month period (November to April) from the previous year was used as a baseline (before Bacillus supplementation) to control for seasonal variations.

[0344] Statistical analysis

[0345] Reproductive indices were evaluated by the PROC MIXED procedure of SAS (version 9.2 SAS Institute, Cary NC). Significance between means was defined at p ≤ 0.05, and trends were indicated at p ≤ 0.1.

[0346] result

[0347] Feeding 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) improved reproductive metrics in the cattle herd as evidenced by fewer services per conception, higher conception and 21-day pregnancy rates.

[0348] Table 10. Effects of treatments on reproductive efficiency indices

[0349]

[0350] Example 6

[0351] Field trials

[0352] The purpose of the field trial was to determine the effects of supplementation with 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 cow health and milk production parameters on a 370-cow Upper Midwestern Healthy Holstein dairy farm with several sporadic cases of hemorrhagic bowel syndrome (HBS).

[0353] Experimental Design and Methods

[0354] Cows received daily Bacillus supplementation at a rate of 15 g of Bacillus product, which corresponds to 7.35 x 10 per head per day on a standard total mixed ration based on corn silage and alfalfa hay. 9 Live Bacillus colony forming units (CFU). Production and health indicators were monitored throughout the herd by Dairy Comp 305 (Valley Agricultural Software, Tulare, CA). Milk production, milk composition and health event data were collected 4 months before Bacillus supplementation (serving as a baseline) and 4 months after the start of Bacillus supplementation. Rumination data were collected 5 months before the start of Bacillus supplementation to serve as a baseline and compared with 5 months after Bacillus supplementation.

[0355] Statistical analysis

[0356] Milk production and milk composition data were analyzed using PROC MIXED of Statistical Analysis System (SAS) software with treatment*month*year and treatment as main effects and days in milk (DIM) as covariate. Health event data were analyzed using PROC GLIMMIX of SAS with main effect of treatment. Rumination data were analyzed by PROC MIXED using treatment as main effect and N as covariate. Significance between means was defined at p≤0.05, and trends were indicated when p≤0.1.

[0357] result

[0358] Feeding 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) for 4 months reduced the incidence of pneumonia in the entire herd.

[0359] Table 11. Effect of treatment on the mean percentage of postpartum cows or the entire herd experiencing pneumonia

[0360]

[0361] Feeding 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) increased daily rumination in the entire herd.

[0362] Table 12. Effects of treatments on mean rumination

[0363]

[0364] Feeding 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) increased milk yield, milk fat percentage, and pounds of milk fat and milk protein, resulting in increased total fat-corrected milk as well as energy-corrected milk across the herd.

[0365] Table 13. Effects of treatments on mean milk yield, percentage of milk components and daily energy-corrected milk yield

[0366]

[0367] Feeding 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) reduced the number of log somatic cell counts present in milk.

[0368] Table 14. Effects of treatments on mean log somatic cell counts.

[0369] <110> Microbiome Discovery Group, LLC United Animal Health, Inc. <120> Microorganisms for animals <130> 60421-299808 <140> PCT / US2019 / 054190 <141> 2019-10-02 <150> 62 / 740,169 <151> 2018-10-02 <160> 3 <170> PatentIn Version 3.5 <210> 1 <211> 10 <212> DNA <213> Artificial sequence <220> <223> Description of artificial sequences: Synthetic primers <400> 1 ggtgcgggaa 10 <210> 2 <211> 10 <212> DNA <213> Artificial sequence <220> <223> Description of artificial sequences: Synthetic primers <400> 2 gtttcgctcc 10 <210> 3 <211> 10 <212> DNA <213> Artificial sequence <220> <223> Description of artificial sequences: Synthetic primers <400> 3 gtagacccgt 10

Claims

1. Use of an additive in preparing a feed composition or in preparing drinking water containing the additive, wherein the feed composition or drinking water is used to improve the reproductive performance of animals, wherein the additive comprises all of the following isolated strains, wherein the isolated strains include: Bacillus strain 86 deposited with NRRL No. B-50944, Bacillus strain 300 deposited with NRRL No. B-50943, Bacillus strain 101 deposited with NRRL No. B-67218, Bacillus strain 235 deposited with NRRL No. B-67219, Bacillus strain 177 deposited with NRRL No. B-67275 and Bacillus strain 102 deposited with NRRL No. B-67276.

2. Use of an additive in preparing a feed composition or in preparing drinking water comprising the additive, wherein the feed composition or drinking water is used in a method for reducing animal mortality and / or reducing animal fasting events, wherein the additive comprises all of the following isolated strains, wherein the isolated strains include: Bacillus strain 86 deposited with NRRL No. B-50944, Bacillus strain 300 deposited with NRRL No. B-50943, Bacillus strain 101 deposited with NRRL No. B-67218, Bacillus strain 235 deposited with NRRL No. B-67219, Bacillus strain 177 deposited with NRRL No. B-67275 and Bacillus strain 102 deposited with NRRL No. B-67276.

3. The use according to claim 1, wherein the animal is selected from poultry species.

4. The use according to claim 1, wherein the animal is selected from the group consisting of porcine species, bovine species, ovine species and equine species.

5. The use according to claim 1, wherein the animal is a porcine species, and the porcine species is selected from the group consisting of growing-finishing pigs and nursery pigs.

6. The use according to claim 1, wherein the animal is a porcine species and the porcine species is a sow.

7. The use according to claim 1, wherein the animal is a porcine species and the porcine species is a breeder pig.

8. The use according to claim 1, wherein the animal is a milk-producing animal.

9. The use according to claim 8, wherein the milk-producing animal is selected from cows or goats.

10. The use according to claim 8, wherein the milk-producing animal is selected from sheep or buffalo.

11. The use according to claim 1, wherein the isolated Bacillus strain produces an enzyme selected from the group consisting of α-galactosidase, protease, lipase, amylase, xylanase, cellulase, and combinations thereof.

12. The use according to claim 1, wherein the feed composition or drinking water additionally comprises another bacterial strain selected from another Bacillus strain, a lactic acid bacteria strain and a combination thereof.

13. The use according to claim 1, wherein each Bacillus strain is present at 1.0 x 10 3 CFU / lb feed composition to 5.0 x 10 12 A dosage of CFU / lb of feed composition is present in the feed composition.

14. The use according to claim 1, wherein each Bacillus strain is present at 0.5 x 10 7 CFU / lb feed composition to 5.0 x 10 7 A dosage of CFU / lb of feed composition is present in the feed composition.

15. Use according to claim 1, wherein the feed composition or drinking water additionally comprises an antibiotic.

16. The use according to claim 15, wherein the antibiotic is selected from erythromycin, levofloxacin, a combination of trimethoprim and sulfamethoxazole, trimethoprim, daptomycin, rifampicin, tylosin, tilmicosin, vancomycin, monensin, lasalocid and lelomycin propionate.

17. The use according to claim 1, wherein the feed composition or drinking water further comprises an enzyme selected from the group consisting of β-galactosidase, protease, lipase, amylase, xylanase, cellulase, phytase and combinations thereof.

18. Use according to claim 17, wherein the enzyme is phytase.

19. Use according to claim 1, wherein the feed composition or drinking water additionally comprises NSPase.

20. The use according to claim 1, wherein the animal is a cow and the feed composition or drinking water comprising the Bacillus strain is administered during lactation.

21. The use according to claim 1, wherein the animal is a cow and the feed composition or drinking water comprising the Bacillus strain is administered during gestation.

22. The use according to claim 1, wherein the feed composition or drinking water comprising the Bacillus strain is administered to the animal daily.

23. The use according to claim 1, wherein the improvement in reproductive performance is selected from the group consisting of a reduction in the number of conception services, an increase in estrus detection, an increase in conception rate, a reduction in miscarriage and an increase in pregnancy rate.

24. A method for improving milk production and / or milk quality of a dairy animal, the method comprising the step of administering to the animal a feed composition or drinking water, wherein the feed composition or drinking water comprises an effective amount of an additive comprising all of the following isolated strains, the isolated strains comprising: Bacillus strain 86 deposited under NRRL No. B-50944, Bacillus strain 300 deposited under NRRL No. B-50943, Bacillus strain 101 deposited under NRRL No. B-67218, Bacillus strain 235 deposited under NRRL No. B-67219, Bacillus strain 177 deposited under NRRL No. B-67275 and Bacillus strain 102 deposited under NRRL No. B-67276, wherein the Bacillus strain improves the milk production and / or milk quality of the animal.

25. Use of a feed composition for the preparation of a medicament for inhibiting a disease or disorder in a milk-producing animal, wherein the disease or disorder is selected from the group consisting of lactogenic fever, ketosis, retained placenta, metritis, pneumonia and ectopic abomasum, the feed composition comprising an effective amount of an additive comprising all of the following isolated strains, the isolated strains comprising: Bacillus strain 86 deposited under NRRL No. B-50944, Bacillus strain 300 deposited under NRRL No. B-50943, Bacillus strain 101 deposited under NRRL No. B-67218, Bacillus strain 235 deposited under NRRL No. B-67219, Bacillus strain 177 deposited under NRRL No. B-67275 and Bacillus strain 102 deposited under NRRL No. B-67276, wherein the Bacillus strain inhibits the disease or disorder in the milk producing animal.

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