Methods for treating high somatic cell counts
The present invention provides methods for treating an animal having a high somatic cell count. The methods include administering compositions 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 treating an animal having a high somatic cell count, the method comprising administering to a milk producing animal having or at risk of having a somatic cell count of at least about 250,000 cells per milliliter of milk an effective amount of a 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 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 siderophore receptor polypeptides have molecular weights of between about 60 kDa and about 100 kDa as determined by separation by sodium dodecyl-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 separation by sodium dodecyl-polyacrylamide gel electrophoresis.
8. The method of claim 1 wherein the animal is a bovine.
9. The method of claim 1 wherein the milk producing animal has or is at risk of having a somatic cell count of at least about 400,000 cells per milliliter of milk.
10. A method for treating an animal having a high somatic cell count, the method comprising administering to a milk producing animal having or at risk of having a somatic cell count of at least about 250,000 cells per milliliter of milk 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.
11. The method of claim 10 wherein the isolated molecules further comprise Iipopolysaccharide at a concentration of no greater than about 10.0 EU/ml.
12. The method of claim 10 wherein the gram negative microbe is an enteropathogen.
13. The method of claim 10 wherein the gram negative microbe is a member of the family Enterobacteriaceae.
14. The method of claim 10 wherein the gram negative microbe is a member of the tribe Escherichieae or Salmonelleae.
15. The method of claim 10 wherein the gram negative microbe is Salmonella spp . or Eschericliia coli.
16. The method of claim 10 wherein the at least two siderophore receptor polypeptides have molecular weights of between about 60 kDa and about 100 kDa as determined by separation by sodium dodecyl-polyacrylamide gel electrophoresis.
17. The method of claim 10 wherein the at least two porins have molecular weights of between about 30 kDa and about 43 kDa as determined by separation by sodium dodecyl-polyacrylamide gel electrophoresis.
18. The method of claim 10 wherein the animal is a bovine.
19. The method of claim 10 wherein the milk producing animal has or is at risk of having a somatic cell count of at least about 400,000 cells per milliliter of milk.