OPTIMIZED DELIVERY OF MICROORGANISMS THROUGH THE WATER MAINS

Claims

1. A composition comprising a) a buffer sufficient to maintain the pH of water at a level of about or greater than about 6.5; and b) one or more direct fed microorganisms (DFM).

2. The composition according to claim 1, additionally containing c) a thickener.

3. The composition according to claim 2, wherein the thickener is xanthan gum.

4. The composition according to any one of claims 1-3, wherein the DFM are freeze-dried or lyophilized.

5. The composition according to claim 4, wherein the DFM contain a cryoprotectant.

6. The composition according to any one of claims 1-3, wherein the composition is hydrated.

7. The composition of any one of claims 1 to 6, wherein the buffer is sufficient to maintain the pH of the water in the water main at a level of about or greater than about 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.0, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, or 9.

0.

8. The composition according to any one of claims 1-7, containing about 1-2000 mM buffer.

9. The composition of claim 8, comprising approximately 320 mM buffer.

10. The composition of claim 7, comprising approximately 0.001-20 mM buffer.

11. The composition of claim 10, comprising approximately 2.5 mM buffer.

12. The composition of any one of claims 3-11, comprising approximately 0.1% w / w xanthan gum.

13. The composition according to any one of claims. 1-12, wherein the buffer comprises one or more buffers selected from the group consisting of potassium phosphate, carbonate, TAPS ([tris(hydroxymethyl)methylamino]propanesulfonic acid), bicine (2-(bis(2-hydroxyethyl)amino)acetic acid), tris (tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol), tricine (N-[tris(hydroxymethyl)methyl]glycine), TAPSO (3-[N-tris(hydroxymethyl)methylamino]-2-hydroxypropanesulfonic acid), HEPES (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid), TES (2-[[1,3-dihydroxy-2-(hydroxymethyl)propan-2-yl]amino]ethanesulfonic acid), MOPS (3-(N-morpholino)propanesulfonic acids), PIPES (piperazine-N,N'-bis(2-ethanesulfonic acid)), cacodylate (dimethylarsenic acid) and MES (2-(N-morpholino)ethanesulfonic acid.

14. The composition according to any one of claims 1-13, wherein the DFM comprises one or more DFM selected from the group consisting of bacteria, algae and fungi.

15. The composition of claim 14, wherein the bacterium comprises a gram-positive bacterium or a gram-negative bacterium.

16. The composition of claim 14, wherein the fungus comprises yeast.

17. The composition according to any one of claims 14 or 15, wherein the bacterium is one or more bacteria selected from the group consisting of Bacillus spp., Bifidobacterium spp., Lactobacillus spp. and Megasphaera spp.

18. The composition of claim 17, wherein the bacterium is one or more bacteria selected from the group consisting of Bacillus subtilis, Bacillus amyloliquefaciens, Bacillus velezensis, Bifidobacterium animalis subspecies lactis, Lactobacillus reuteri, Lactobacillus acidophilus and Megasphaera elsdenii.

19. The composition of any one of claims 6-18, wherein the DFM exhibit reduced settling and / or improved survival in a hydrated composition compared to identical DFM that are not in a hydrated composition containing a buffer sufficient to maintain the pH of the water at a level of about or greater than about 6.

5.

20. The composition according to any one of claims 1-19, wherein the water is selected from the group consisting of water from a municipal source, water from an underground source, surface water, and collected rainwater.

21. The composition according to claim 20, wherein chloramine or another chlorine-based antimicrobial agent is added to the water.

22. The composition according to claim 21, further comprising d) means for inactivating chloramine or chlorine present in the water.

23. The composition according to any one of claims 1-22, further comprising e) a wetting agent and / or a dispersing agent.

24. The composition of any one of claims 1-23, wherein the temperature of the composition is less than about 15°C.

25. A method for producing a composition for maximizing the delivery of live cells of direct fed microorganisms (DFM) through a water main, comprising mixing i) a buffer sufficient to maintain the pH of water at a level of about or greater than about 6.5; and ii) one or more direct fed microorganisms (DFM) with water.

26. The method according to claim 25, wherein the DFM is freeze-dried or lyophilized.

27. The method according to claim 25, wherein the DFM contain a cryoprotectant.

28. The method according to any one of paragraphs 25-27, wherein the water is selected from the group consisting of water from a municipal source, water from an underground source, surface water and collected rainwater.

29. The method according to any one of paragraphs 25-28, further comprising mixing iii) the thickener with water.

30. The method according to any one of claims 25-29, wherein the mixing provides a final buffer concentration of approximately 1-2000 mM.

31. The method of claim 30, wherein the mixing provides a final buffer concentration of approximately 320 mM.

32. The method according to any one of paragraphs 29-31, providing approximately 0.1% w / w thickener.

33. The method of any one of paragraphs 29-32, wherein the buffer is sufficient to maintain the pH of the water in the water main at a level of about or greater than about 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.0, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, or 9.

0.

34. The method of any one of claims 25 to 33, wherein the buffer is one or more buffers selected from the group consisting of potassium phosphate, carbonate, TAPS ([tris(hydroxymethyl)methylamino]propanesulfonic acid), bicine (2-(bis(2-hydroxyethyl)amino)acetic acid), tris (tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol), tricine (N-[tris(hydroxymethyl)methyl]glycine), TAPSO (3-[N-tris(hydroxymethyl)methylamino]-2-hydroxypropanesulfonic acid), HEPES (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid), TES (2-[[1,3-dihydroxy-2-(hydroxymethyl)propan-2-yl]amino]ethanesulfonic acid), MOPS (3-(N-morpholino)propanesulfonic acid), PIPES (piperazine-N,N'-bis(2-ethanesulfonic acid)), cacodylate (dimethylarsenic acid) and MES (2-(N-morpholino)ethanesulfonic acid.

35. The method according to any one of claims 25-34, wherein the DFM is one or more DFM selected from the group consisting of bacteria and fungi.

36. The method according to claim 35, wherein the bacterium is a gram-positive bacterium or a gram-negative bacterium.

37. The method according to claim 35, wherein the fungus is yeast.

38. The method according to any one of claims 35 or 36, wherein the bacterium is one or more bacteria selected from the group consisting of Bacillus spp., Bifidobacterium spp., Lactobacillus spp. and Megasphaera spp.

39. The method according to claim 38, wherein the bacterium is one or more bacteria selected from the group consisting of Bacillus subtilis, Bacillus amyloliquefaciens, Bacillus velezensis, Bifidobacterium animalis subspecies lactis, Lactobacillus reuteri, Lactobacillus acidophilus and Megasphaera elsdenii.

40. The method according to any one of paragraphs 25-39, wherein chloramine or another chlorine-based antimicrobial agent is added to the water.

41. The method of claim 40, further comprising mixing iv) means for inactivating chloramine or chlorine present in water with water.

42. The method according to any one of paragraphs 25-41, further comprising mixing v) a wetting agent and / or a dispersing agent with water.

43. The method according to any one of claims 25-42, wherein the temperature of the composition is less than about 15°C.

44. A composition obtained by the method according to any of paragraphs 25-43.

45. A method of delivering one or more direct fed microorganisms (DFM) to a subject, comprising administering a composition according to any one of claims 1-24 or 44 via a water main, carried out within a certain distance, where the DFM are delivered to the subject if the subject drinks from the water main.

46. ​​The method of claim 45, wherein the DFM exhibit reduced settling and / or survival in the water main compared to identical DFM that are not incorporated into the compositions of any one of claims 1-24 or 44.

47. The method according to claim 45 or 46, wherein the water main comprises nipple drinkers.

48. The method according to any one of paragraphs 45-47, wherein the distance is from about 20 meters to about 200 meters.

49. The method of claim 48, wherein the water main further comprises from approximately 20 to 100 meters of pipeline in which there are no nipple drinkers.

50. The method according to any one of paragraphs 45-49, wherein the subject is a poultry, pig or ruminant animal.

51. The method according to claim 50, wherein the poultry is a chicken or a turkey.

52. The method according to claim 50 or 51, wherein the chicken is a layer or broiler.

53. The method of claim 50, wherein the pig is a piglet, a growing pig, a fattening pig, or a sow.

54. The method according to claim 50, wherein the ruminant animal is beef cattle, dairy cattle or calves.

55. A kit comprising a) a buffer sufficient to maintain the pH of water at or above about 6.5; and b) one or more direct fed microorganisms (DFM).

56. The kit according to item 55, additionally containing c) a thickener.

57. The kit according to claim 56, wherein the thickener is xanthan gum.

58. The kit of any one of paragraphs 55-57, wherein the buffer is sufficient to maintain the pH of the water in the water main at a level of about or greater than about 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.0, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, or 9.

0.

59. A kit according to any one of paragraphs 55-58, containing 3-500 g of buffer.

60. Recruitment according to any of paragraphs. 55-59, wherein the buffer comprises one or more buffers selected from the group consisting of potassium phosphate, carbonate, TAPS ([tris(hydroxymethyl)methylamino]propanesulfonic acid), bicine (2-(bis(2-hydroxyethyl)amino)acetic acid), tris (tris(hydroxymethyl)aminomethane or 2-amino-2-(hydroxymethyl)propane-1,3-diol), tricine (N-[tris(hydroxymethyl)methyl]glycine), TAPSO (3-[N-tris(hydroxymethyl)methylamino]-2-hydroxypropanesulfonic acid), HEPES (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid), TES (2-[[1,3-dihydroxy-2-(hydroxymethyl)propan-2-yl]amino]ethanesulfonic acid), MOPS (3-(N-morpholino)propanesulfonic acids), PIPES (piperazine-N,N'-bis(2-ethanesulfonic acid)), cacodylate (dimethylarsenic acid) and MES (2-(N-morpholino)ethanesulfonic acid.

61. The kit according to any one of claims 55-60, wherein the DFM comprises one or more DFM selected from the group consisting of bacteria, algae and fungi.

62. The kit according to claim 61, wherein the bacterium comprises a gram-positive bacterium or a gram-negative bacterium.

63. The kit according to claim 61, wherein the fungus comprises yeast.

64. The kit according to any one of claims 61 or 62, wherein the bacterium is one or more bacteria selected from the group consisting of Bacillus spp., Bifidobacterium spp., Lactobacillus spp. and Megasphaera spp.

65. The kit of claim 64, wherein the bacterium is one or more bacteria selected from the group consisting of Bacillus subtilis, Bacillus amyloliquefaciens, Bacillus velezensis, Bifidobacterium animalis subspecies lactis, Lactobacillus reuteri, Lactobacillus acidophilus and Megasphaera elsdenii.

66. A kit according to any one of paragraphs 55-65, further comprising d) means for inactivating chloramine or chlorine present in water.

67. A kit according to any one of claims 55-66, wherein the components of the kit are packaged in sachets, water-soluble tablets, water-soluble sachets (e.g. powder distributed in cold water-soluble PVA film bags), or water-soluble (gel) capsules (e.g. pullulan capsules).

68. A kit according to any one of paragraphs 66-67, further comprising e) written instructions for combining the components of the kit with water for delivery via a water main.

69. The kit according to any one of claims 55-68, wherein the DFM are freeze-dried or lyophilized.

70. The kit according to item 69, wherein the DFM additionally contain a cryoprotectant.

71. A kit according to any one of paragraphs 55-70, additionally containing f) a wetting and / or dispersing agent.