Compositions and methods for inhibition of pathogenic bacterial growth
Compositions for methods of preventing or reducing pathogenic bacterial growth, proliferation, and/or colonization are described containing one or more types of non-pathogenic bacteria to be introduced into the gastrointestinal tract and effectively compete with pathogenic bacteria for monomeric or polymeric carbohydrate nutrients, and/or amino acid nutrients, and/or vitamin nutrients.
1. A composition comprising a population of bacteria, an excipient, and an enteric coating, the composition formulated for oral administration to a mammalian subject, wherein the bacteria consist of spore forming bacteria comprising at least a first and a second spore forming species of bacteria, wherein the first spore forming species of bacteria is Clostridium bolteae comprising a 16S sequence having at least 97% sequence identity to SEQ ID No: 559, and the second spore forming species of bacteria is Clostridium orbiscindens comprising a 16S sequence having at least 97% sequence identity to SEQ ID No: 609, wherein the first and second spore forming species of bacteria are: (i) isolated, (ii) not identical, and (iii) independently capable of proliferating in a medium comprising a nutrient and having a threshold concentration of the nutrient below a concentration of the nutrient required for Clostridium difficile proliferation in the medium.
2. The composition of claim 1 , wherein the nutrient comprises a carbohydrate nutrient.
3. The composition of claim 1 , wherein the threshold concentration is 90% or less of the concentration of the nutrient required for Clostridium difficile proliferation.
4. The composition of claim 1 , wherein the threshold concentration is 50% or less of the concentration of the nutrient required for Clostridium difficile proliferation.
5. The composition of claim 1 , wherein the threshold concentration is 10%, 5%, or 1% of the concentration of the nutrient required for Clostridium difficile proliferation.
6. The composition of claim 1 , wherein the nutrient comprises a vitamin nutrient.
7. The composition of claim 1 , wherein the first and second spore forming species of bacteria are independently capable of proliferating at a rate at least 10% greater than Clostridium difficile proliferation rate in the medium, wherein the nutrient comprises one or more carbohydrate nutrients.
8. A method of treating or preventing Clostridium difficile infection in a mammalian subject, the method comprising administering to the subject an effective amount of the composition of claim 1 , wherein the effective amount prevents or reduces Clostridium difficile growth, proliferation, and/or colonization in the subject.
9. The composition of claim 1 , wherein the excipient is selected from the group consisting of: a buffering agent, a preservative, a stabilizer, a binder, a compaction agent, a lubricant, a dispersion enhancer, a disintegration agent, a flavoring agent, a sweetener, and a coloring agent.
10. The composition of claim 1 , wherein the excipient comprises glycerol.
11. The composition of claim 1 , wherein proliferation in the medium is determined by inoculating into the medium at least one bacteria selected from the group consisting of the first spore forming species of bacteria, the second spore forming species of bacteria and Clostridium difficile , culturing the at least one bacteria in the medium for a time period, measuring a titer of the at least one bacteria in the medium at several points over the time period, and calculating a growth rate as a difference in the titer over the time period.
12. The composition of claim 11 , wherein the medium consists of 0.5% (wt/vol) of a test carbohydrate, 0.125 mg/l biotin, 1 mg/l pyridoxine and pantothenate, 75 mg/l histidine, glycine and tryptophan, 150 mg/l arginine, methionine and threonine, 225 mg/l valine and isoleucine, 300 mg/l leucine, 400 mg/l cysteine, and 450 mg/l proline; and the titer is measured as an OD600; and the growth rate is calculated ln(X/X o )/T, where X is the OD600 value during a linear portion of growth and T is time (hours).
13. The composition of claim 1 , wherein the population of bacteria consists of spores.
14. The composition of claim 1 , wherein the spore forming bacteria further comprises a third spore forming species of bacteria, wherein the third spore forming species of bacteria is Clostridium scindens comprising a 16S sequence having at least 97% sequence identity to SEQ ID No: 623.
15. The composition of claim 1 , wherein the spore forming bacteria further comprises a third spore forming species of bacteria, wherein the third spore forming species of bacteria is Clostridium symbiosum comprising a 16S sequence having at least 97% sequence identity to SEQ ID No: 652.
16. The composition of claim 1 , wherein the spore forming bacteria further comprises a third and a fourth spore forming species of bacteria, wherein the third spore forming species of bacteria is Clostridium scindens comprising a 16S sequence having at least 97% sequence identity to SEQ ID No: 623 and the fourth spore forming species of bacteria is Clostridium symbiosum comprising a 16S sequence having at least 97% sequence identity to SEQ ID No: 652.