Methods for reducing fecal shedding
The present invention provides methods for reducing shedding in an animal. Generally, the method includes administcriirn to an animal a composition including siderophore receptor polypeptides and porins from gram negative microbes, and preferably, lipopolysaccarhide at a concentration of no greater than about 10.0 endotoxin units per milliliter.
1. A method for reducing fecal shedding of a microbe in an animal's intestinal tract, the method comprising administering to an animal an effective amount of a composition comprising:
at least two siderophore receptor polypeptides (SRPs) isolated from a gram negative microbe;
at least two porins isolated from the gram negative microbe;
lipopolysaccharide at a concentration of no greater than about 10.0 EU/ml; and
a pharmaceutically acceptable carrier.
2. The method of claim 1 wherein the gram negative microbe is an enteropathogen.
3. The method of claim 1 wherein the gram negative microbe is a member of the family Enterobacteriaceae.
4. The method of claim 1 wherein the gram negative microbe is a member of the tribe Escherichieae or Salmonelleae.
5. The method of claim 1 wherein the gram negative microbe is Salmonella spp. or Escherichia coli.
6. The method of claim 1 wherein the at least two SRPs have molecular weights of between about 60 kDa and about 100 kDa as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis.
7. The method of claim 1 wherein the at least two porins have molecular weights of between about 30 kDa and about 43 kDa as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis.
8. The method of claim 1 wherein the animal is an avian, a bovine, a caprine, a porcine, or an ovine.
9. The method of claim 1 wherein the microbe present in the animal's intestinal tract is a gram negative microbe or a gram positive microbe.
10. The method of claim 1 wherein the fecal shedding of the microbe is decreased when compared to the focal shedding of the microbe before receiving the composition.
11. The method of claim 1 wherein the microbe being shed by the animal is Salmonella spp. or E, coli.
12. The method of claim 1 wherein the gram negative microbe is Salmonella spp., and wherein each of the at least two SRPs have a molecular weight selected from the group consisting of between about 87 kDa and about 91 kDa, between about 82 kDa and about 86 kDa, and between about 69 kDa and about 75 kDa as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis.
13. A method for reducing fecal shedding of a microbe in an animal's intestinal tract, the method comprising administering to an animial an effective amount of a first composition comprising:
at least two siderophore receptor polypeptides isolated from a gram negative microbe;
at least two porins isolated from the gram negative microbe;
lipopolysaccharide; and
a pharmaceutically acceptable carrier, wherein the concentration of lipopolysaccharide present in the first composition is no greater than the concentration of lipopolysaccharide in a second composition comprising:
the at least two siderophore receptor polypeptides isolated from the gram negative microbe;
the at least two porins isolated from the gram negative microbe;
lipopolysaccharide;
wherein the second composition is produced by a process comprising:
providing the gram negative microbe;
disrupting the gram negative microbe in a buffer;
solubilizing the disrupted gram negative microbe for greater than about 24 hours in a solution comprising sarcosine to result in solubilized and insoluble cellular material, wherein a ratio of the sarcosine to gram weight of disrupted gram negative microbe is between about 0.8 gram sarcosine per about 4.5 grams of disrupted gram negative microbe and about 1.2 grams sarcosine per about 4.5 grams of disrupted gram negative microbe; and
isolating molecules of the gram negative microbe, wherein the isolated molecules comprise the at least two siderophore receptor polypeptides, the at least two porins, and lipopolysaccharide.
14. The method of claim 13 wherein the concentration of lipopolysaccharide in the first composition is no greater than about 10.0 EU/ml.
15. The method of claim 13 wherein the gram negative microbe is an enteropathogen.
16. The method of claim 13 wherein the gram negative microbe is a member of the family Enterobacteriaceae.
17. The method of claim 13 wherein the gram negative microbe is a member of the tribe Escherichieae or Salmonelleae.
18. The method of claim 13 wherein the gram negative microbe is Salmonella spp. or Escherichia coli.
19. The method of claim 13 wherein the at least two SRPs have molecular weights of between about 60 kDa and about 100 kDa as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis.
20. The method of claim 13 wherein the at least two porins have molecular weights of between about 30 kDa and about 43 kDa as determined by sodium dodecyl sulfate-polyacrylarmide gel electrophoresis.
21. The method of claim 13 wherein the animal is an avian, a bovine, a caprine, a porcine, or an ovine.
22. The method of claim 13 wherein the microbe present in the animal's intestinal tract is a gram negative microbe or a gram positive microbe.
23. The method of claim 13 wherein the fecal shedding of the microbe is decreased when compared to the fecal shedding of the microbe before receiving the composition.
24. The method of claim 1 wherein the microbe being shed by the animal is Salmonella spp. or E coli.
25. The method of claim 1 wherein the gram negative microbe is Salmonella spp., and wherein each of the at least two SRPs have a molecular weight selected from the group consisting of between about 87 kDa and about 91 kDa, between about 82 kDa and about 86 kDa, and between about 69 kDa and about 75 kDa, as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis.