Process for the manufacture of pathogen inhibiting bacteria

• US 9,901,605 B2
• Filed: 08/25/2017
• Issued: 02/27/2018
• Est. Priority Date: 02/01/2010
• Status: Active Grant

• Current Assignee: MicroBios Inc.
• Original Assignee: MicroBios Inc.
• Inventors: Garner, Matthew Ryan, Flint, Joseph F.
• Primary Examiner(s): NAVARRO, ALBERT MARK
• Application Number:
  US15/686,971
• Publication Number:
  US 20170348364A1
• Time in Patent Office: 186 Days
• CPC Class Codes: A23K 10/18 of live microorganisms
• CPC Class Codes: A23K 50/10 for ruminants
• CPC Class Codes: A23K 50/20 for horses
• CPC Class Codes: A23K 50/30 for swines
• CPC Class Codes: A23K 50/75 for poultry
• CPC Class Codes: A23K 50/80 for aquatic animals, e.g. fish, crustaceans or molluscs
• CPC Class Codes: A23V 2400/181 Salivarius
• CPC Class Codes: A61K 35/747 Lactobacilli, e.g. L. acidophilus or L. brevis
• CPC Class Codes: C12N 1/04 Preserving or maintaining viable microorganisms immobilised microorganisms C12N11/00
• CPC Class Codes: C12N 1/20 Bacteria; Culture media therefor
• CPC Class Codes: C12N 1/205 Bacterial isolates
• CPC Class Codes: C12R 2001/23 Lactobacillus acidophilus

Abstract

Processes to produce microorganisms that can be incorporated into a microbial-based product that results in high viable cell yields and shelf-stable products are disclosed. These microbial-based products are useful for inhibiting pathogenic growth and as a food additive. A preferred microorganism is a lactic acid producing bacteria. In one embodiment, the process comprises inoculating a lactobacillus fermentation medium with lactic acid producing bacterial cells, harvesting the lactic acid producing bacterial cells at least partially during the log phases, concentrating the lactic acid producing bacterial cells, and preserving the lactic acid producing bacterial cells at a concentration of at least 5×10⁹ cfu/ml.

Claims

1. A method of manufacturing a composition comprising live bacterial cells, the method comprising:

inoculating a bacterial fermentation medium with live Lactobacillus animalis cells strain MB101 having ATCC Accession Number PTA-121710; and

harvesting the live Lactobacillus animalis cells during at least one logarithmic phase of growth.

2. The method as recited in claim 1, further comprising:

harvesting the live Lactobacillus animalis cells strain MB101 having ATCC Accession Number PTA-121710 at least partially during at least one of mid-log phase of growth and late-log phase of growth.

3. The method as recited in claim 2, further comprising:

harvesting the live Lactobacillus animalis cells strain MB101 having ATCC Accession Number PTA-121710 during mid-log phase of growth.

4. The method as recited in claim 2, further comprising:

harvesting the live Lactobacillus animalis cells strain MB101 having ATCC Accession Number PTA-121710 during late-log phase of growth.

5. The method as recited in claim 1, further comprising:

harvesting the live Lactobacillus animalis cells strain MB101 having ATCC Accession Number PTA-121710 between mid-log phase of growth and late-log phase of growth.

6. The method as recited in claim 1, further comprising:

determining, prior to harvesting, that a cell count of the live Lactobacillus animalis cells strain MB101 having ATCC Accession Number PTA-121710 is at least 1×

10⁶ cells/ml.

7. The method as recited in claim 1, further comprising:

harvesting the live Lactobacillus animalis cells strain MB101 having ATCC Accession Number PTA-121710 at least partially during mid-log phase of growth and at least partially during another log phase of growth.

8. The method as recited in claim 1, further comprising concentrating the live Lactobacillus animalis cells strain MB101 having ATCC Accession Number PTA-121710 to a live cell count of at least 5×

10⁹ cells/ml.

9. The method as recited in claim 1, further comprising:

harvesting the live Lactobacillus animalis cells strain MB101 having ATCC Accession Number PTA-121710 when the live cells are substantially round-shaped.

10. The method as recited in claim 1, further comprising:

harvesting the live Lactobacillus animalis cells strain MB101 having ATCC Accession Number PTA-121710 when the live cells are substantially spherically shaped.

11. The method as recited in claim 1, further comprising:

harvesting the live Lactobacillus animalis cells strain MB101 having ATCC Accession Number PTA-121710 before the live cells grow to an elongate shape.

12. A method of manufacturing a composition comprising live bacterial cells, the method comprising:

inoculating a bacterial fermentation medium with live Lactobacillus animalis cells strain MB102 having ATCC Accession Number PTA-121711; and

harvesting the live Lactobacillus animalis cells during at least one logarithmic phase of growth.

13. The method as recited in claim 12, further comprising:

harvesting the live Lactobacillus animalis cells strain MB102 having ATCC Accession Number PTA-121711 at least partially during at least one of mid-log phase of growth and late-log phase of growth.

14. The method as recited in claim 13, further comprising:

harvesting the live Lactobacillus animalis cells strain MB102 having ATCC Accession Number PTA-121711 during mid-log phase of growth.

15. The method as recited in claim 13, further comprising:

harvesting the live Lactobacillus animalis cells strain MB102 having ATCC Accession Number PTA-121711 during late-log phase of growth.

16. The method as recited in claim 12, further comprising:

determining, prior to harvesting, that a cell count of the live Lactobacillus animalis cells strain MB101 having ATCC Accession Number PTA-121710 is at least 1×

10⁶ cells/ml.

17. The method as recited in claim 12, further comprising concentrating the live Lactobacillus animalis cells strain MB102 having ATCC Accession Number PTA-121711 to a live cell count of at least 5×

10⁹ cells/ml.

18. The method as recited in claim 12, further comprising:

harvesting the live Lactobacillus animalis cells strain MB102 having ATCC Accession Number PTA-121711 when the live cells are substantially round-shaped.

19. The method as recited in claim 12, further comprising:

harvesting the live Lactobacillus animalis cells strain MB102 having ATCC Accession Number PTA-121711 when the live cells are substantially spherically shaped.

20. The method as recited in claim 12, further comprising:

harvesting the live Lactobacillus animalis cells strain MB102 having ATCC Accession Number PTA-121711 before the live cells grow to an elongate shape.